Polynucleotides encoding STE20-related protein kinases and methods of use

ABSTRACT

The present invention relates to the nucleic acid molecules encoding an STE20-related family of novel protein kinases, ZC1, ZC2, ZC3, ZC4, STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, KHS2, SULU1, SULU3, GEK2, PAK4 and PAK5, segments and domains thereof, as well as various methods useful for the diagnosis and treatment of various kinase-related diseases and conditions. Mammalian nucleic acid molecules encoding these kinases are particularly disclosed, and more specifically human sources of these nucleic acids are disclosed.

RELATED APPLICATIONS

The present application claims priority to U.S. Provisional Patent Application Ser. No. 60/081,784 by Plowman and Martinez, entitled STE20-Related Protein kinases, filed Apr. 14, 1998, hereby incorporated by reference herein in its entirety, including any drawings, tables, or figures.

The instant application contains a “lengthy” Sequence Listing which has been submitted via triplicate CD-R in lieu of a printed paper copy, and is hereby incorporated by reference in its entirety. Said CD-R are labeled “CRE”, “Copy 1” and “Copy 2”, respectively, and each contains only one identical 329 Kb file (38602329.APP).

FIELD OF THE INVENTION

The present invention relates to novel kinase polypeptides, nucleotide sequences encoding the novel kinase polypeptides, as well as various products and methods useful for the diagnosis and treatment of various kinase-related diseases and conditions.

BACKGROUND OF THE INVENTION

The following description of the background of the invention is provided to aid in understanding the invention, but is not admitted to be or to describe prior art to the invention.

Cellular signal transduction is a fundamental mechanism whereby external stimuli that regulate diverse cellular processes are relayed to the interior of cells. One of the key biochemical mechanisms of signal transduction involves the reversible phosphorylation of proteins, which enables regulation of the activity of mature proteins by altering their structure and function.

The best characterized protein kinases in eukaryotes phosphorylate proteins on the hydroxyl moiety of serine, threonine and tyrosine residues. These kinases largely fall into two groups, those specific for phosphorylating serines and threonines, and those specific for phosphorylating tyrosines. Some kinases, referred to as “dual specificity” kinases, are able to phosphorylate on tyrosine as well as serine/threonine residues.

Protein kinases can also be characterized by their location within the cell. Some kinases are transmembrane receptor-type proteins capable of directly altering their catalytic activity in response to the external environment such as the binding of a ligand. Others are non-receptor-type proteins lacking any transmembrane domain. They can be found in a variety of cellular compartments from the inner surface of the cell membrane to the nucleus.

Many kinases are involved in regulatory cascades wherein their substrates may include other kinases whose activities are regulated by their phosphorylation state. Ultimately the activity of some downstream effector is modulated by phosphorylation resulting from activation of such a pathway.

Protein kinases are one of the largest families of eukaryotic proteins with several hundred known members. These proteins share a 250-300 amino acid domain that can be subdivided into 12 distinct subdomains that comprise the common catalytic core structure. These conserved protein motifs have recently been exploited using PCR-based cloning strategies leading to a significant expansion of the known kinases.

Multiple alignment of the sequences in the catalytic domain of protein kinases and subsequent parsimony analysis permits the segregation of related kinases into distinct branches or subfamilies including: tyrosine kinases, cyclic-nucleotide-dependent kinases, calcium/calmodulin kinases, cyclin-dependent kinases and MAP-kinases, serine-threonine kinase receptors, and several other less defined subfamilies.

SUMMARY OF THE INVENTION

Through the use of a targeted PCR cloning strategy and of a “motif extraction” bioinformatics script, mammalian members of the STE20-kinase family have been identified as part of the present invention. Multiple alignment and parsimony analysis of the catalytic domain of all of these STE20-family members reveals that these proteins cluster into 9 distinct subgroups. Classification in this manner has proven highly accurate not only in predicting motifs present in the remaining non-catalytic portion of each protein, but also in their regulation, substrates, and signaling pathways. The present invention includes the partial or complete sequence of new members of the STE20-family, their classification, predicted or deduced protein structure, and a strategy for elucidating their biologic and therapeutic relevance.

Thus, a first aspect of the invention features an isolated, enriched, or purified nucleic acid molecule encoding a kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5.

By “isolated” in reference to nucleic acid is meant a polymer of nucleotides conjugated to each other, including DNA and RNA, that is isolated from a natural source or that is synthesized. The isolated nucleic acid of the present invention is unique in the sense that it is not found in a pure or separated state in nature. Use of the term “isolated” indicates that a naturally occurring sequence has been removed from its normal cellular (i.e., chromosomal) environment. Thus, the sequence may be in a cell-free solution or placed in a different cellular environment. The term does not imply that the sequence is the only nucleotide chain present, but that it is essentially free (about 90-95% pure at least) of non-nucleotide material naturally associated with it, and thus is distinguished from isolated chromosomes.

By the use of the term “enriched” in reference to nucleic acid is meant that the specific DNA or RNA sequence constitutes a significantly higher fraction (2-5 fold) of the total DNA or RNA present in the cells or solution of interest than in normal or diseased cells or in the cells from which the sequence was taken. This could be caused by a person by preferential reduction in the amount of other DNA or RNA present, or by a preferential increase in the amount of the specific DNA or RNA sequence, or by a combination of the two. However, it should be noted that enriched does not imply that there are no other DNA or RNA sequences present, just that the relative amount of the sequence of interest has been significantly increased. The term “significant” is used to indicate that the level of increase is useful to the person making such an increase, and generally means an increase relative to other nucleic acids of about at least 2 fold, more preferably at least 5 to 10 fold or even more. The term also does not imply that there is no DNA or RNA from other sources. The other source DNA may, for example, comprise DNA from a yeast or bacterial genome, or a cloning vector such as pUC19. This term distinguishes from naturally occurring events, such as viral infection, or tumor type growths, in which the level of one mRNA may be naturally increased relative to other species of mRNA. That is, the term is meant to cover only those situations in which a person has intervened to elevate the proportion of the desired nucleic acid.

It is also advantageous for some purposes that a nucleotide sequence be in purified form. The term “purified” in reference to nucleic acid does not require absolute purity (such as a homogeneous preparation). Instead, it represents an indication that the sequence is relatively more pure than in the natural environment (compared to the natural level this level should be at least 2-5 fold greater, e.g., in terms of mg/mL). Individual clones isolated from a cDNA library may be purified to electrophoretic homogeneity. The claimed DNA molecules obtained from these clones could be obtained directly from total DNA or from total RNA. The cDNA clones are not naturally occurring, but rather are preferably obtained via manipulation of a partially purified naturally occurring substance (messenger RNA). The construction of a cDNA library from mRNA involves the creation of a synthetic substance (cDNA) and pure individual cDNA clones can be isolated from the synthetic library by clonal selection of the cells carrying the cDNA library. Thus, the process which includes the construction of a cDNA library from mRNA and isolation of distinct cDNA clones yields an approximately 10⁶-fold purification of the native message. Thus, purification of at least one order of magnitude, preferably two or three orders, and more preferably four or five orders of magnitude is expressly contemplated.

By a “kinase polypeptide” is meant 32 (preferably 40, more preferably 45, most preferably 55) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, or the corresponding full-length amino acid sequence; 250 (preferably 255, more preferably 260, most preferably 270) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, or SEQ ID NO:105, or the corresponding full-length amino acid sequence; 27 (preferably 30, more preferably 40, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:18; 16 (preferably 20, more preferably 25, most preferably 35) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, or SEQ ID NO:103 or the corresponding full-length amino acid sequence; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99, 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101, or the corresponding full-length amino acid sequence; 78 (preferably 80, more preferably 85, most preferably 90) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:107 or functional derivatives thereof as described herein. For sequences for which the full-length sequence is not given, the remaining sequences can be determined using methods well-known to those in the art and are intended to be included in the invention. In certain aspects, polypeptides of 100, 200, 300 or more amino acids are preferred. The kinase polypeptide can be encoded by a full-length nucleic acid sequence or any portion of the full-length nucleic acid sequence, so long as a functional activity of the polypeptide is retained, not to include fragments containing only amino acids 1-22 of SEQ ID NO:13 or only amino acids 1-33 of SEQ ID NO:107.

The amino acid sequence will be substantially similar to the sequence shown in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequence, or fragments thereof, not to include fragments consisting only of the amino acid sequences 1-22 of SEQ ID NO:13 or 1-33 of SEQ ID NO:107. A sequence that is substantially similar to the sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107 will preferably have at least 90% identity (more preferably at least 95% and most preferably 99-100%) to the sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107.

By “identity” is meant a property of sequences that measures their similarity or relationship. Identity is measured by dividing the number of identical residues by the total number of residues and gaps and multiplying the product by 100. “Gaps” are spaces in an alignment that are the result of additons or deletions of amino acids. Thus, two copies of exactly the same sequence have 100% identity, but sequences that are less highly conserved, and have deletions, additions, or replacements, may have a lower degree of identity. Those skilled in the art will recognize that several computer programs are available for determining sequence identity using standard parameters, for example Blast (Altschul, et al. (1997) Nucleic Acids Res. 25:3389-3402), Blast2 (Altschul, et al. (1990) J. mol. biol. 215:403-410), and Smith-Waterman (Smith, et al. (1981) J. Mol. Biol. 147:195-197).

In preferred embodiments, the invention features isolated, enriched, or purified nucleic acid molecules encoding a kinase polypeptide comprising a nucleotide sequence that: (a) encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107; (b) is the complement of the nucleotide sequence of (a); (c) hybridizes under highly stringent conditions to the nucleotide molecule of (a) and encodes a naturally occurring kinase polypeptide; (d) encodes a kinase polypeptide having the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO: 103, SEQ ID NO:105, or SEQ ID NO:107, except that it lacks one or more, but not all, of the following segments of amino acid residues: 1-21, 22-274, or 275-416 of SEQ ID NO:5, 1-31, 32-308, 309-489 or 490-516 of SEQ ID NO:6, 1-178 or 179-414 of SEQ ID NO:7, 1-22, 23-289, 290-526, 527-640, 641-896, or 897-1239 of SEQ ID NO:13, 1-255, 256-442, 443-626, 627-954, or 955-1297 of SEQ ID NO:14, 1-255, 256-476, 477-680, 681-983, or 984-1326 of SEQ ID NO:15, 1-13, 14-273, 274-346, 347-534, or 535-894 of SEQ ID NO:18, 1-21, 22-277, 278-427, 428-637, 638-751, or 752-898 of SEQ ID NO:22, 1-66, 67-215, 216-425, 426-539, 540-786, or 787-887 of SEQ ID NO:23, 1-25, 26-273, 274-422, 423-632, or 633-748 of SEQ ID NO:24, 1-51, 52-224, 225-393, 394-658, or 659-681 of SEQ ID NO:29, 1-25, 26-281, 284-430, 431-640, 641-754, 755-901, or 902-1001 of SEQ ID NO:31, 1-10, 11-321, or 322-373 of SEQ ID NO:97, 1-57, 58-369, or 370-418 of SEQ ID NO:99, 1-52, 53-173, 174-307, 308-572, or 573-591 of SEQ ID NO:103, 1-24, 25-289, 290-397, 398-628, 629-872, or 873-1227 of SEQ ID NO:105, or 1-33, 34-294, 295-337, 338-472, 473-724, or 725-968 of SEQ ID NO:107; (e) is the complement of the nucleotide sequence of (d); (f) encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31; SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107 from amino acid residues 1-21, 22-274, or 275-416 of SEQ ID NO:5, 1-31, 32-308, 309-489, or 490-516 of SEQ ID NO:6, 1-178 or 179-414 of SEQ ID NO:7, 23-289, 290-526, 527-640, 641-896, or 897-1239 of SEQ ID NO:13, 1-255, 256-442, 443-626, 627-954, or 955-1297 of SEQ ID NO:14, 1-255, 256-476, 477-680, 681-983, or 984-1326 of SEQ ID NO:15, 1-13, 14-273, 274-346, 347-534, or 535-894 of SEQ ID NO:18, 1-21, 22-277, 278-427, 428-637, 638-751, or 752-898 of SEQ ID NO:22, 1-66, 67-215, 216-425, 426-539, 540-786, or 787-887 of SEQ ID NO:23, 1-25, 26-273, 274-422, 423-632, or 633-748 of SEQ ID NO:24, 1-51, 52-224, 225-393, 394-658, or 659-681 of SEQ ID NO:29, 1-25, 26-281, 282-430, 431-640, 641-754, 755-901, or 902-1001 of SEQ ID NO:31, 1-10, 11-321, or 322-373 of SEQ ID NO:97, 1-57, 58-369, or 370-418 of SEQ ID NO:99, 1-52, 53-173, 174-307, 308-572, or 573-591 of SEQ ID NO:103, 1-24, 25-289, 290-397, 398-628, 629-872, or 873-1227 of SEQ ID NO:105,or 1-33, 34-294, 295-337, 338-472, 473-724, or 725-968 of SEQ ID NO:107; (g) is the complement of the nucleotide sequence of (f);(h) encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, except that it lacks one or more of the domains selected from the group consisting of a N-terminal domain, a catalytic domain, a C-terminal domain, a coiled-coil structure region, a proline-rich region, a spacer region, an insert, and a C-terminal tail; or (i) is the complement of the nucleotide sequence of (h).

The term “complement” refers to two nucleotides that can form multiple favorable interactions with one another. For example, adenine is complementary to thymine as they can form two hydrogen bonds. Similarly, guanine and cytosine are complementary since they can form three hydrogen bonds. A nucleotide sequence is the complement of another nucleotide sequence if all of the nucleotides of the first sequence are complementary to all of the nucleotides of the second sequence.

The term “domain” refers to a region of a polypeptide which contains a particular function. For instance, N-terminal or C-terminal domains of signal transduction proteins can serve functions including, but not limited to, binding molecules that localize the signal transduction molecule to different regions of the cell or binding other signaling molecules directly responsible for propagating a particular cellular signal. Some domains can be expressed separately from the rest of the protein and function by themselves, while others must remain part of the intact protein to retain function. The latter are termed functional regions of proteins and also relate to domains.

The term “N-terminal domain” refers to the extracatalytic region located between the initiator methionine and the catalytic domain of the protein kinase. The N-terminal domain can be identified following a Smith-Waterman alignment of the protein sequence against the non-redundant protein database to define the N-terminal boundary of the catalytic domain. Depending on its length, the N-terminal domain may or may not play a regulatory role in kinase function. An example of a protein kinase whose N-terminal domain has been shown to play a regulatory role is PAK65, which contains a CRIB motif used for Cdc42 and rac binding (Burbelo, P.D. et al. (1995) J. Biol. Chem. 270, 29071-290740).

The N-terminal domain spans amino acid residues 1-21 of the sequence set forth in SEQ ID NO:5, amino acid residues 1-31 of the sequence set forth in SEQ ID NO:6, amino acid residues 1-22 of the sequence set forth in SEQ ID NO:13, amino acid residues 1-13 of the sequence set forth in SEQ ID NO:18, amino acid residues 1-21 of the sequence set forth in SEQ ID NO:22, amino acid residues 1-25 of the sequence set forth in SEQ ID NO:24, amino acid residues 1-51 of the sequence set forth in SEQ ID NO:29, amino acid residues 1-25 of the sequence set forth in SEQ ID NO:31, amino acid residues 1-57 of the sequence set forth in SEQ ID NO:99, amino acid residues 1-52 of the sequence set forth in SEQ ID NO:103, amino acid residues 1-24 of the sequence set forth in SEQ ID NO:105, or amino acid residues 1-33 of the sequence set forth in SEQ ID NO:107.

The term “catalytic domain” refers to a region of the protein kinase that is typically 25-300 amino acids long and is responsible for carrying out the phosphate transfer reaction from a high-energy phosphate donor molecule such as ATP or GTP to itself (autophosphorylation) or to other proteins (exogenous phosphorylation). The catalytic domain of protein kinases is made up of 12 subdomains that contain highly conserved amino acid residues, and are responsible for proper polypeptide folding and for catalysis. The catalytic domain can be identified following a Smith-Waterman alignment of the protein sequence against the non-redundant protein database.

The catalytic domain spans amino acid residues 22-274 of the sequence set forth in SEQ ID NO:5, residues 32-308 of the sequence set forth in SEQ ID NO:6, residues 1-178 of the sequence set forth in SEQ ID NO:7, residues 23-289 of the sequence set forth in SEQ ID NO:13, residues 1-255 of the sequence set forth in SEQ ID NO:14, residues 1-255 of the sequence set forth in SEQ ID NO:15, residues 14-273 of the sequence set forth in SEQ ID NO:18, residues 22-277 of the sequence set forth in SEQ ID NO:22, residues 1-66 of the sequence set forth in SEQ ID NO:23, residues 26-273 of the sequence set forth in SEQ ID NO:24, residues 394-658 of the sequence set forth in SEQ ID NO:29, residues 26-281 of the sequence set forth in SEQ ID NO:31, residues 1-278 of the sequence set forth in SEQ ID NO:97, residues 58-369 of the sequence set forth in SEQ ID NO:99, residues 1-103 of the sequence set forth in SEQ ID NO:101, residues 308-572 of the sequence set forth in SEQ ID NO:103, residues 25-289 of the sequence set forth in SEQ ID NO:105, or residues 34-294 of the sequence set forth in SEQ ID NO:107.

The term “catalytic activity”, as used herein, defines the rate at which a kinase catalytic domain phosphorylates a substrate. Catalytic activity can be measured, for example, by determining the amount of a substrate converted to a phosphorylated product as a function of time. Catalytic activity can be measured by methods of the invention by holding time constant and determining the concentration of a phosphorylated substrate after a fixed period of time. Phosphorylation of a substrate occurs at the active-site of a protein kinase. The active-site is normally a cavity in which the substrate binds to the protein kinase and is phosphorylated.

The term “substrate” as used herein refers to a molecule phosphorylated by a kinase of the invention. Kinases phosphorylate substrates on serine/threonine or tyrosine amino acids. The molecule may be another protein or a polypeptide.

The term “C-terminal domain” refers to the region located between the catalytic domain or the last (located closest to the C-terminus) functional domain and the carboxy-terminal amino acid residue of the protein kinase. By “functional” domain is meant any region of the polypeptide that may play a regulatory or catalytic role as predicted from amino acid sequence homology to other proteins or by the presence of amino acid sequences that may give rise to specific structural conformations (i.e. coiled-coils). The C-terminal domain can be identified by using a Smith-Waterman alignment of the protein sequence against the non-redundant protein database to define the C-terminal boundary of the catalytic domain or of any functional C-terminal extracatalytic domain. Depending on its length and amino acid composition, the C-terminal domain may or may not play a regulatory role in kinase function. An example of a protein kinase whose C-terminal domain may play a regulatory role is PAK3 which contains a heterotrimeric Gb subunit-binding site near its C-terminus (Leeuw, T. et al (1998) Nature, 391, 191-195).

The C-terminal domain spans amino acid residues 275-416 of the sequence set forth in SEQ ID NO:5, residues 309-489 of the sequence set forth in SEQ ID NO:6, residues 179-414 of the sequence set forth in SEQ ID NO:7, residues 897-1239 of the sequence set forth in SEQ ID NO:13, residues 955-1297 of the sequence set forth in SEQ ID NO:14, residues 984-1326 of the sequence set forth in SEQ ID NO:15, residues 535-894 of the sequence set forth in SEQ ID NO:18, residues 752-898 of the sequence set forth in SEQ ID NO:22, residues 279-330 of the sequence set forth in SEQ ID NO:97, residues 370-418 of the sequence set forth in SEQ ID NO:99, or residues 873-1227 of the sequence set forth in SEQ ID NO:105.

The term “signal transduction pathway” refers to the molecules that propagate an extracellular signal through the cell membrane to become an intracellular signal. This signal can then stimulate a cellular response. The polypeptide molecules involved in signal transduction processes are typically receptor and non-receptor protein tyrosine kinases, receptor and non-receptor protein phosphatases, SRC homology 2 and 3 domains, phosphotyrosine binding proteins (SRC homology 2 (SH2) and phosphotyrosine binding (PTB and PH) domain containing proteins), proline-rich binding proteins (SH3 domain containing proteins), nucleotide exchange factors, and transcription factors.

The term “coiled-coil structure region” as used herein, refers to a polypeptide sequence that has a high probability of adopting a coiled-coil structure as predicted by computer algorithms such as COILS (Lupas, A. (1996) Meth. Enzymology 266:513-525). Coiled-coils are formed by two or three amphipathic α-helices in parallel. Coiled-coils can bind to coiled-coil domains of other polypeptides resulting in homo- or heterodimers (Lupas, A. (1991) Science 252:1162-1164). Coiled-coil-dependent oligomerization has been shown to be necessary for protein function including catalytic activity of serine/threonine kinases (Roe, J. et al. (1997) J. Biol. Chem. 272:5838-5845).

The coiled-coil structure region spans amino acid residues 290-526 of the sequence set forth in SEQ ID NO:13, residues 256-442 of the sequence set forth in SEQ ID NO:14, residues 256-476 of the sequence set forth in SEQ ID NO:15, residues 428-637 of the sequence set forth in SEQ ID NO:22, residues 216-425 or 540-786 of the sequence set forth in SEQ ID NO:23, residues 423-632 of the sequence set forth in SEQ ID NO:24, residues 431-640 or 755-901 of the sequence set forth in SEQ ID NO:31, residues 291-398 or 629-668 of the sequence set forth in SEQ ID NO:105, or residues 473-724 or 725-968 of the sequence set forth in SEQ ID NO:107.

The term “proline-rich region” as used herein, refers to a region of a protein kinase whose proline content over a given amino acid length is higher than the average content of this amino acid found in proteins (i.e., >10%). Proline-rich regions are easily discernable by visual inspection of amino acid sequences and quantitated by standard computer sequence analysis programs such as the DNAStar program EditSeq. Proline-rich regions have been demonstrated to participate in regulatory protein—protein interactions. Among these interactions, those that are most relevant to this invention involve the “PxxP” (SEQ ID NO: 148) proline rich motif found in certain protein kinases (i.e., human PAM) and the SH3 domain of the adaptor molecule Nck (Galisteo, M. L. et al. (1996) J. Biol. Chem. 271:20997-21000). Other regulatory interactions involving “PxxP” (SEQ ID NO: 148) proline-rich motifs include the WW domain (Sudol, M. (1996) Prog. Biochys. Mol. Bio. 65:113-132).

The proline-rich region spans amino acid residues 527-640 of the sequence set forth in SEQ ID NO:13, residues 443-626 of the sequence set forth in SEQ ID NO:14, residues 477-680 of the sequence set forth in SEQ ID NO:15, residues 347-534 of the sequence set forth in SEQ ID NO:18,residues 398-628 of the sequence set forth in SEQ ID NO:105, or residues 338-472 of the sequence set forth in SEQ ID NO:107.

The term “spacer region” as used herein, refers to a region of the protein kinase located between predicted functional domains. The spacer region has no detectable homology to any amino acid sequence in the database, and can be identified by using a Smith-Waterman alignment of the protein sequence against the non-redundant protein database to define the C- and N-terminal boundaries of the flanking functional domains. Spacer regions may or may not play a fundamental role in protein kinase function. Precedence for the regulatory role of spacer regions in kinase function is provided by the role of the src kinase spacer in inter-domain interactions (Xu, W. et al. (1997) Nature 385:595-602).

The spacer region spans amino acid residues 641-896 of the sequence set forth in SEQ ID NO:13, residues 627-954 of the sequence set forth in SEQ ID NO:14, residues 681-983 of the sequence set forth in SEQ ID NO:15, residues 274-346 of the sequence set forth in SEQ ID NO:18, residues 278-427 or 638-751 of the sequence set forth in SEQ ID NO:22, residues 67-215 or 426-539 of the sequence set forth in SEQ ID NO:23, residues 274-422 or 633-748 of the sequence set forth in SEQ ID NO:24, residues 225-393 of the sequence set forth in SEQ ID NO:29, residues 282-430 or 641-754 of the sequence set forth in SEQ ID NO:31, residues 174-307 of the sequence set forth in SEQ ID NO:103, residues 669-872 of the sequence set forth in SEQ ID NO:105, or residues 295-337 of the sequence set forth in SEQ ID NO:107.

The term “insert” as used herein refers to a portion of a protein kinase that is absent from a close homolog. Inserts may or may not by the product alternative splicing of exons. Inserts can be identified by using a Smith-Waterman sequence alignment of the protein sequence against the non-redundant protein database, or by means of a multiple sequence alignment of homologous sequences using the DNAStar program Megalign. Inserts may play a functional role by presenting a new interface for protein—protein interactions, or by interfering with such interactions. Inserts span amino acid residues 52-224 of the sequence set forth in SEQ ID NO:29 or residues 53-173 of the sequence set forth in SEQ ID NO:103.

The term “C-terminal tail” as used herein, refers to a C-terminal domain of a protein kinase, that by homology extends or protrudes past the C-terminal amino acid of its closest homolog. C-terminal tails can be identified by using a Smith-Waterman sequence alignment of the protein sequence against the non-redundant protein database, or by means of a multiple sequence alignment of homologous sequences using the DNAStar program Megalign. Depending on its length, a C-terminal tail may or may not play a regulatory role in kinase function.

The C-terminal tail spans amino acid residues 490-516 of the sequence set forth in SEQ ID NO:6, residues 787-887 of the sequence set forth in SEQ ID NO:23, residues 659-681 of the sequence set forth in SEQ ID NO:29, residues 994-1093 of the sequence set forth in SEQ ID NO:31, or residues 573-591 of the sequence set forth in SEQ ID NO:103.

Various low or high stringency hybridization conditions may be used depending upon the specificity and selectivity desired. These conditions are well-known to those skilled in the art. Under stringent hybridization conditions only highly complementary nucleic acid sequences hybridize. Preferably, such conditions prevent hybridization of nucleic acids having more than 1 or 2 mismatches out of 20 contiguous nucleotides, more preferably, such conditions prevent hybridization of nucleic acids having more than 1 or 2 mismatches out of 50 contiguous nucleotides, most preferably, such conditions prevent hybridization of nucleic acids having more than 1 or 2 mismatches out of 100 contiguous nucleotides. In some instances, the conditions may prevent hybridization of nucleic acids having more than 5 mismatches in the full-length sequence.

By stringent hybridization assay conditions is meant hybridization assay conditions at least as stringent as the following: hybridization in 50% formamide, 5× SSC, 50 mM NaH₂PO₄, pH 6.8, 0.5% SDS, 0.1 mg/mL sonicated salmon sperm DNA, and 5× Denhart solution at 42° C. overnight; washing with 2× SSC, 0.1% SDS at 45° C.; and washing with 0.2× SSC, 0.1% SDS at 45° C. Under some of the most stringent hybridization assay conditions, the second wash can be done with 0.1× SSC at a temperature up to 70° C. (Berger et al. (1987) Guide to Molecular Cloning Techniques pg 421, hereby incorporated by reference herein including any figures, tables, or drawings.). However, other applications may require the use of conditions falling between these sets of conditions. Methods of determining the conditions required to achieve desired hybridizations are well-known to those with ordinary skill in the art, and are based on several factors, including but not limited to, the sequences to be hybridized and the samples to be tested.

In other preferred embodiments, the invention features isolated, enriched, or purified nucleic acid molecules encoding kinase polypeptides, further comprising a vector or promoter effective to initiate transcription in a host cell. The invention also features recombinant nucleic acid, preferably in a cell or an organism. The recombinant nucleic acid may contain a sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102,SEQ ID NO:104, or SEQ ID NO:106, or a functional derivative thereof and a vector or a promoter effective to initiate transcription in a host cell. The recombinant nucleic acid can alternatively contain a transcriptional initiation region functional in a cell, a sequence complementary to an RNA sequence encoding a kinase polypeptide and a transcriptional termination region functional in a cell. Specific vectors and host cell combinations are discussed herein.

The term “vector” relates to a single or double-stranded circular nucleic acid molecule that can be transfected into cells and replicated within or independently of a cell genome. A circular double-stranded nucleic acid molecule can be cut and thereby linearized upon treatment with restriction enzymes. An assortment of nucleic acid vectors, restriction enzymes, and the knowledge of the nucleotide sequences cut by restriction enzymes are readily available to those skilled in the art. A nucleic acid molecule encoding a kinase can be inserted into a vector by cutting the vector with restriction enzymes and ligating the two pieces together.

The term “transfecting” defines a number of methods to insert a nucleic acid vector or other nucleic acid molecules into a cellular organism. These methods involve a variety of techniques, such as treating the cells with high concentrations of salt, an electric field, detergent, or DMSO to render the outer membrane or wall of the cells permeable to nucleic acid molecules of interest or use of various viral transduction strategies.

The term “promoter” as used herein, refers to nucleic acid sequence needed for gene sequence expression. Promoter regions vary from organism to organism, but are well known to persons skilled in the art for different organisms. For example, in prokaryotes, the promoter region contains both the promoter (which directs the initiation of RNA transcription) as well as the DNA sequences which, when transcribed into RNA, will signal synthesis initiation. Such regions will normally include those 5′-non-coding sequences involved with initiation of transcription and translation, such as the TATA box, capping sequence, CAAT sequence, and the like.

In preferred embodiments, the isolated nucleic acid comprises, consists essentially of, or consists of a nucleic acid sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100 SEQ ID NO:102, SEQ ID NO:104, or SEQ ID NO:106, or the corresponding full-length sequence, encodes the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequence, a functional derivative thereof, or at least 40, 45, 50, 60, 100, 200, or 300 contiguous amino acids of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, or of the corresponding full-length amino acid sequence; at least 250, 255, 275, 300, or 400 contiguous amino acids of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, or of the corresponding full-length amino acid sequence; at least 27, 30, 35, 40, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:18; at least 16, 25, 35, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, or SEQ ID NO:103, or of the corresponding full-length amino acid sequence; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99, or the corresponding full-length amino acid sequence; 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101, or the corresponding full-length amino acid sequence; or at least 80, 85, 90, 100, 200, or 300 contiguous amino acids of SEQ ID NO:107, or functional derivatives thereof. The kinase polypeptides, selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5, comprise, consist essentially of, or consist of at least at least 40, 45, 50, 60, 100, 200, or 300 contiguous amino acids of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7; at least 250, 255, 275, 300, or 400 contiguous amino acids of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, or SEQ ID NO:105; at least 27, 30, 35, 40, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:18; at least 35, 40, 45, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31 or SEQ ID NO:103; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99; 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101; or at least 80, 85, 90, 100, 200, or 300 contiguous amino acids of SEQ ID NO:107, or the corresponding full-length sequences or derivatives thereof. The nucleic acid may be isolated from a natural source by cDNA cloning or by subtractive hybridization. The natural source may be mammalian, preferably human, blood, semen, or tissue, and the nucleic acid may be synthesized by the triester method or by using an automated DNA synthesizer.

The term “mammal” refers preferably to such organisms as mice, rats, rabbits, guinea pigs, sheep, and goats, more preferably to cats, dogs, monkeys, and apes, and most preferably to humans.

In yet other preferred embodiments, the nucleic acid is a conserved or unique region, for example those useful for: the design of hybridization probes to facilitate identification and cloning of additional polypeptides, the design of PCR probes to facilitate cloning of additional polypeptides, obtaining antibodies to polypeptide regions, and designing antisense oligonucleotides.

By “conserved nucleic acid regions”, are meant regions present on two or more nucleic acids encoding a kinase polypeptide, to which a particular nucleic acid sequence can hybridize under lower stringency conditions. Examples of lower stringency conditions suitable for screening for nucleic acid encoding kinase polypeptides are provided in Abe, et al. (J. Biol. Chem. 19:13361-13368, 1992), hereby incorporated by reference herein in its entirety, including any drawings, figures, or tables. Preferably, conserved regions differ by no more than 5 out of 20 nucleotides, even more preferably 2 out of 20 nucleotides or most preferably 1 out of 20 nucleotides.

By “unique nucleic acid region” is meant a sequence present in a nucleic acid coding for a kinase polypeptide that is not present in a sequence coding for any other naturally occurring polypeptide. Such regions preferably encode 32 (preferably 40, more preferably 45, most preferably 55) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, or the corresponding full-length amino acid sequence; 250 (preferably 255, more preferably 260, most preferably 270) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:15, or SEQ ID NO:105, or the corresponding full-length amino acid sequence; 27 (preferably 30, more preferably 40, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:18; 16 (preferably 20, more preferably 25, most preferably 35) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, or SEQ ID NO:103, or the corresponding full-length amino acid sequence; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99, 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101, or the corresponding full-length amino acid sequence; or 78 (preferably 80, more preferably 85, most preferably 90) or more contiguous amino acids set forth in the amino acid sequence SEQ ID NO:107, or functional derivatives thereof. In particular, a unique nucleic acid region is preferably of mammalian origin.

A second aspect of the invention features a nucleic acid probe for the detection of nucleic acid encoding a kinase polypeptide in a sample, wherein said polypeptide is selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. Preferably, the nucleic acid probe encodes a kinase polypeptide that is a fragment of the protein encoded by the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequences, not to include fragments consisting only of amino acids 1-22 of SEQ ID NO:13 or amino acids 1-33 of SEQ ID NO:107. The nucleic acid probe contains a nucleotide base sequence that will hybridize to a sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104, or SEQ ID NO:106, or the corresponding full-length sequence, or a functional derivative thereof.

In preferred embodiments, the nucleic acid probe hybridizes to nucleic acid encoding at least 6, 12, 75, 90, 105, 120, 150, 200, 250, 300 or 350 contiguous amino acids of the sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31 SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequence, or functional derivatives thereof.

Methods for using the probes include detecting the presence or amount of kinase RNA in a sample by contacting the sample with a nucleic acid probe under conditions such that hybridization occurs and detecting the presence or amount of the probe bound to kinase RNA. The nucleic acid duplex formed between the probe and a nucleic acid sequence coding for a kinase polypeptide may be used in the identification of the sequence of the nucleic acid detected (Nelson et al., in Nonisotopic DNA Probe Techniques, Academic Press, San Diego, Kricka, ed., p. 275, 1992, hereby incorporated by reference herein in its entirety, including any drawings, figures, or tables). Kits for performing such methods may be constructed to include a container means having disposed therein a nucleic acid probe.

In a third aspect, the invention describes a recombinant cell or tissue comprising a nucleic acid molecule encoding a kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. In such cells, the nucleic acid may be under the control of the genomic regulatory elements, or may be under the control of exogenous regulatory elements including an exogenous promoter. By “exogenous” it is meant a promoter that is not normally coupled in vivo transcriptionally to the coding sequence for the kinase polypeptides.

The polypeptide is preferably a fragment of the protein encoded by the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequence, not to include fragments consisting only of amino acids 1-22 of SEQ ID NO:13 or amino acids 1-33 of SEQ ID NO:107. By “fragment,” is meant an amino acid sequence present in a kinase polypeptide. Preferably, such a sequence comprises at least 32, 45, 50, 60, 100, 200, or 300 contiguous amino acids of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, or of the corresponding full-length amino acid sequence; at least 250, 255, 275, 300, or 400 contiguous amino acids of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, OR SEQ ID NO:105, or of the corresponding full-length amino acid sequence; at least 27, 30, 35, 40, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:18; at least 16, 25, 35, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31 or SEQ ID NO:103, or of the corresponding full-length amino acid sequence; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99, 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101; at least 78, 85, 90, 100, 200, or 300 contiguous amino acids of SEQ ID NO:107, or the corresponding full-length amino acid sequence; or a functional derivative thereof.

In a fourth aspect, the invention features an isolated, enriched, or purified kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5.

By “isolated” in reference to a polypeptide is meant a polymer of amino acids (2 or more amino acids) conjugated to each other, including polypeptides that are isolated from a natural source or that are synthesized. The isolated polypeptides of the present invention are unique in the sense that they are not found in a pure or separated state in nature. Use of the term “isolated” indicates that a naturally occurring sequence has been removed from its normal cellular environment. Thus, the sequence may be in a cell-free solution or placed in a different cellular environment. The term does not imply that the sequence is the only amino acid chain present, but that it is essentially free (about 90-95% pure at least) of non-amino acid material naturally associated with it.

By the use of the term “enriched” in reference to a polypeptide is meant that the specific amino acid sequence constitutes a significantly higher fraction (2-5 fold) of the total amino acid sequences present in the cells or solution of interest than in normal or diseased cells or in the cells from which the sequence was taken. This could be caused by a person by preferential reduction in the amount of other amino acid sequences present, or by a preferential increase in the amount of the specific amino acid sequence of interest, or by a combination of the two. However, it should be noted that enriched does not imply that there are no other amino acid sequences present, just that the relative amount of the sequence of interest has been significantly increased. The term significant here is used to indicate that the level of increase is useful to the person making such an increase, and generally means an increase relative to other amino acid sequences of about at least 2-fold, more preferably at least 5- to 10-fold or even more. The term also does not imply that there is no amino acid sequence from other sources. The other source of amino acid sequences may, for example, comprise amino acid sequence encoded by a yeast or bacterial genome, or a cloning vector such as pUC19. The term is meant to cover only those situations in which man has intervened to increase the proportion of the desired amino acid sequence.

It is also advantageous for some purposes that an amino acid sequence be in purified form. The term “purified” in reference to a polypeptide does not require absolute purity (such as a homogeneous preparation); instead, it represents an indication that the sequence is relatively purer than in the natural environment. Compared to the natural level this level should be at least 2-5 fold greater (e.g., in terms of mg/mL). Purification of at least one order of magnitude, preferably two or three orders, and more preferably four or five orders of magnitude is expressly contemplated. The substance is preferably free of contamination at a functionally significant level, for example 90%, 95%, or 99% pure.

In preferred embodiments, the kinase polypeptide is a fragment of the protein encoded by the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequences, not to include fragments consisting only of amino acids 1-22 of SEQ ID NO:13 or amino acids 1-33 of SEQ ID NO:107. Preferably, the kinase polypeptide contains at least 32, 45, 50, 60, 100, 200, or 300 contiguous amino acids of SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7, or the corresponding full-length amino acid sequence; at least 250, 255, 275, 300, or 400 contiguous amino acids of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, or SEQ ID NO:105, or the corresponding full-length amino acid sequence; at least 27, 30, 35, 40, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:18; at least 16, 25, 35, 50, 100, 200, or 300 contiguous amino acids of SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, or SEQ ID NO:103, or the corresponding full-length amino acid sequence; 6 (preferably 10, more preferably 15, most preferably 25) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:99, 22 (preferably 30, more preferably 35, most preferably 45) or more contiguous amino acids set forth in the amino acid sequence of SEQ ID NO:101, or the corresponding full-length amino acid sequence; or at least 78, 85, 90, 100, 200, or 300 contiguous amino acids of SEQ ID NO:107, or a functional derivative thereof.

In preferred embodiments, the kinase polypeptide comprises an amino acid sequence having (a) the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107; (b) the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, except that it lacks one or more, but not all, of the following segments of amino acid residues: 1-21, 22-274, or 275-416 of SEQ ID NO:5, 1-31, 32-308, 309-489 or 490-516 of SEQ ID NO:6, 1-178 or 179-414 of SEQ ID NO:7, 1-22, 23-289, 290-526, 527-640, 641-896, or 897-1239 of SEQ ID NO:13, 1-255, 256-442, 443-626, 627-954, or 955-1297 of SEQ ID NO:14, 1-255, 256-476, 477-680, 681-983, or 984-1326 of SEQ ID NO:15, 1-13, 14-273, 274-346, 347-534, or 535-894 of SEQ ID NO:18, 1-21, 22-277, 278-427, 428-637, 638-751, or 752-898 of SEQ ID NO:22, 1-66, 67-215, 216-425, 426-539, 540-786, or 787-887 of SEQ ID NO:23, 1-25, 26-273, 274-422, 423-632, or 633-748 of SEQ ID NO:24, 1-51, 52-224, 225-393, 394-658, or 659-681 of SEQ ID NO:29, 1-25, 26-281, 282-430, 431-640, 641-754, 755-901, or 902-1001 of SEQ ID NO:31, 1-10, 11-321, or 322-373 of SEQ ID NO:97, 1-57, 58-369, or 370-418 of SEQ ID NO:99, 1-52, 53-173, 174-307, 308-572, or 573-591 of SEQ ID NO:103, 1-24, 25-289, 290-397, 398-628, 629-668, 669-872, or 873-1227 of SEQ ID NO:105, or 1-33, 34-294, 295-337, 338-472, 473-724, or 725-968 of SEQ ID NO:107; (c) the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107 from amino acid residues 1-21, 22-274, or 275-416 of SEQ ID NO:5, 1-31, 32-308, 309-489, or 490-516 of SEQ ID NO:6, 1-178 or 179-414 of SEQ ID NO:7, 23-289, 290-526, 527-640, 641-896, or 897-1239 of SEQ ID NO:13, 1-255, 256-442, 443-626, 627-954, or 955-1297 of SEQ ID NO:14, 1-255, 256-476, 477-680, 681-983, or 984-1326 of SEQ ID NO:15, 1-13, 14-273, 274-346, 347-534, or 535-894 of SEQ ID NO:18, 1-21, 22-277, 278-427, 428-637, 638-751, or 752-898 of SEQ ID NO:22, 1-66, 67-215, 216-425, 426-539, 540-786, or 787-887 of SEQ ID NO:23, 1-25, 26-273, 274-422, 423-632, or 633-748 of SEQ ID NO:24, 1-51, 52-224, 225-393, 394-658, or 659-681 of SEQ ID NO:29, 1-25, 26-273, 274-422, 423-632, 633-746, 747-993, or 994-1093 of SEQ ID NO:31, 1-10, 11-321, or 322-373 of SEQ ID NO:97, 1-57, 58-369, or 370-418 of SEQ ID NO:99, 1-52, 53-173, 174-307, 308-572, or 573-591 of SEQ ID NO:103, 1-24, 25-289, 290-397, 398-628, 629-668, 669-872, or 873-1227 of SEQ ID NO:105, or 1-33, 34-294, 295-337, 338-472, 473-724, or 725-968 of SEQ ID NO:107; or (d) the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, except that it lacks one or more, but not all, of the domains selected from the group consisting of a C-terminal domain, a catalytic domain, an N-terminal domain, a spacer region, a proline-rich region, a coiled-coil structure region, an insert, and a C-terminal tail.

The polypeptide can be isolated from a natural source by methods well-known in the art. The natural source may be mammalian, preferably human, blood, semen, or tissue, and the polypeptide may be synthesized using an automated polypeptide synthesizer. The isolated, enriched, or purified kinase polypeptide is preferably: a STLK2, STLK3, STLK4, STLK5, STLK6, or STLK7 polypeptide; a ZC1, ZC2, ZC3, or ZC4 polypeptide; a KHS2 polypeptide; a SULU1 or SULU3 polypeptide; a GEK2 polypeptide; or a PAK4 or PAK5 polypeptide.

In some embodiments the invention includes a recombinant kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. By “recombinant kinase polypeptide” is meant a polypeptide produced by recombinant DNA techniques such that it is distinct from a naturally occurring polypeptide either in its location (e.g., present in a different cell or tissue than found in nature), purity or structure. Generally, such a recombinant polypeptide will be present in a cell in an amount different from that normally observed in nature.

In a fifth aspect, the invention features an antibody (e.g., a monoclonal or polyclonal antibody) having specific binding affinity to a kinase polypeptide or a kinase polypeptide domain or fragment where the polypeptide is selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. By “specific binding affinity” is meant that the antibody binds to the target kinase polypeptide with greater affinity than it binds to other polypeptides under specified conditions. Antibodies or antibody fragments are polypeptides that contain regions that can bind other polypeptides. The term “specific binding affinity” describes an antibody that binds to a kinase polypeptide with greater affinity than it binds to other polypeptides under specified conditions.

The term “polyclonal” refers to antibodies that are heterogenous populations of antibody molecules derived from the sera of animals immunized with an antigen or an antigenic functional derivative thereof. For the production of polyclonal antibodies, various host animals may be immunized by injection with the antigen. Various adjuvants may be used to increase the immunological response, depending on the host species.

“Monoclonal antibodies” are substantially homogenous populations of antibodies to a particular antigen. They may be obtained by any technique which provides for the production of antibody molecules by continuous cell lines in culture. Monoclonal antibodies may be obtained by methods known to those skilled in the art (Kohler et al., Nature 256:495-497, 1975, and U.S. Pat. No. 4,376,110, both of which are hereby incorporated by reference herein in their entirety including any figures, tables, or drawings).

The term “antibody fragment” refers to a portion of an antibody, often the hyper variable region and portions of the surrounding heavy and light chains, that displays specific binding affinity for a particular molecule. A hyper variable region is a portion of an antibody that physically binds to the polypeptide target.

Antibodies or antibody fragments having specific binding affinity to a kinase polypeptide of the invention may be used in methods for detecting the presence and/or amount of kinase polypeptide in a sample by probing the sample with the antibody under conditions suitable for kinase-antibody immunocomplex formation and detecting the presence and/or amount of the antibody conjugated to the kinase polypeptide. Diagnostic kits for performing such methods may be constructed to include antibodies or antibody fragments specific for the kinase as well as a conjugate of a binding partner of the antibodies or the antibodies themselves.

An antibody or antibody fragment with specific binding affinity to a kinase polypeptide of the invention can be isolated, enriched, or purified from a prokaryotic or eukaryotic organism. Routine methods known to those skilled in the art enable production of antibodies or antibody fragments, in both prokaryotic and eukaryotic organisms. Purification, enrichment, and isolation of antibodies, which are polypeptide molecules, are described above.

Antibodies having specific binding affinity to a kinase polypeptide of the invention may be used in methods for detecting the presence and/or amount of kinase polypeptide in a sample by contacting the sample with the antibody under conditions such that an immunocomplex forms and detecting the presence and/or amount of the antibody conjugated to the kinase polypeptide. Diagnostic kits for performing such methods may be constructed to include a first container containing the antibody and a second container having a conjugate of a binding partner of the antibody and a label, such as, for example, a radioisotope. The diagnostic kit may also include notification of an FDA approved use and instructions therefor.

In a sixth aspect, the invention features a hybridoma which produces an antibody having specific binding affinity to a kinase polypeptide or a kinase polypeptide domain, where the polypeptide is selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. By “hybridoma” is meant an immortalized cell line that is capable of secreting an antibody, for example an antibody to a kinase of the invention. In preferred embodiments, the antibody to the kinase comprises a sequence of amino acids that is able to specifically bind a kinase polypeptide of the invention.

In a seventh aspect, the invention features a kinase polypeptide binding agent able to bind to a kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK6, STLK7, STLK5, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. The binding agent is preferably a purified antibody that recognizes an epitope present on a kinase polypeptide of the invention. Other binding agents include molecules that bind to kinase polypeptides and analogous molecules that bind to a kinase polypeptide. Such binding agents may be identified by using assays that measure kinase binding partner activity, such as those that measure PDGFR activity.

The invention also features a method for screening for human cells containing a kinase polypeptide of the invention or an equivalent sequence. The method involves identifying the novel polypeptide in human cells using techniques that are routine and standard in the art, such as those described herein for identifying the kinases of the invention (e.g., cloning, Southern or Northern blot analysis, in situ hybridization, PCR amplification, etc.).

In an eighth aspect, the invention features methods for identifying a substance that modulates kinase activity comprising the steps of: (a) contacting a kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5 with a test substance; (b) measuring the activity of said polypeptide; and (c) determining whether said substance modulates the activity of said polypeptide.

The term “modulates” refers to the ability of a compound to alter the function of a kinase of the invention. A modulator preferably activates or inhibits the activity of a kinase of the invention depending on the concentration of the compound exposed to the kinase.

The term “activates” refers to increasing the cellular activity of the kinase. The term inhibit refers to decreasing the cellular activity of the kinase. Kinase activity is preferably the interaction with a natural binding partner.

The term “modulates” also refers to altering the function of kinases of the invention by increasing or decreasing the probability that a complex forms between the kinase and a natural binding partner. A modulator preferably increases the probability that such a complex forms between the kinase and the natural binding partner, more preferably increases or decreases the probability that a complex forms between the kinase and the natural binding partner depending on the concentration of the compound exposed to the kinase, and most preferably decreases the probability that a complex forms between the kinase and the natural binding partner.

The term “complex” refers to an assembly of at least two molecules bound to one another. Signal transduction complexes often contain at least two protein molecules bound to one another. For instance, a protein tyrosine receptor protein kinase, GRB2, SOS, RAF, and RAS assemble to form a signal transduction complex in response to a mitogenic ligand.

The term “natural binding partner” refers to polypeptides, lipids, small molecules, or nucleic acids that bind to kinases in cells. A change in the interaction between a kinase and a natural binding partner can manifest itself as an increased or decreased probability that the interaction forms, or an increased or decreased concentration of kinase/natural binding partner complex.

The term “contacting” as used herein refers to mixing a solution comprising the test compound with a liquid medium bathing the cells of the methods. The solution comprising the compound may also comprise another component, such as dimethyl sulfoxide (DMSO), which facilitates the uptake of the test compound or compounds into the cells of the methods. The solution comprising the test compound may be added to the medium bathing the cells by utilizing a delivery apparatus, such as a pipet-based device or syringe-based device.

In a ninth aspect, the invention features methods for identifying a substance that modulates kinase activity in a cell comprising the steps of: (a) expressing a kinase polypeptide in a cell, wherein said polypeptide is selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5; (b) adding a test substance to said cell; and (c) monitoring a change in cell phenotype or the interaction between said polypeptide and a natural binding partner.

The term “expressing” as used herein refers to the production of kinases of the invention from a nucleic acid vector containing kinase genes within a cell. The nucleic acid vector is transfected into cells using well known techniques in the art as described herein.

In a tenth aspect, the invention provides methods for treating a disease by administering to a patient in need of such treatment a substance that modulates the activity of a kinase selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5. Preferably, the disease is selected from the group consisting of immune-related diseases and disorders, organ transplantation, myocardial infarction, cardiovascular disease, stroke, renal failure, oxidative stress-related neurodegenerative disorders, and cancer. Most preferably, the immune-related diseases and disorders include, but are not limited to, rheumatoid arthritis, artherosclerosis, and autoimmune disorders.

In preferred embodiments, the invention provides methods for treating or preventing a disease or disorder by administering to a patient in need of such treatment a substance that modulates the activity of a kinase polypeptide selected from the group consisting of ZC1, ZC2, ZC3, ZC4, KHS2, PAK4, and PAK5. Preferably, the disease or disorder is selected from the group consisting of rheumatoid arthritis, artherosclerosis, autoimmune disorders, and organ transplantation. The invention also features methods of treating or preventing a disease or disorder by administering to a patient in need of such treatment a substance that modulates the activity of a kinase polypeptide selected from the group consisting of STLK1, STLK2, STLK3, STLK4, STLK5, STLK6, and STLK7. Preferably the disease or disorder is selected from the group consisting of immune-related diseases and disorders, myocardial infarction, cardiomyopathies, stroke, renal failure, and oxidative stress-related neurodegenerative disorders. Most preferably, the immune-related diseases and disorders are selected from the group consisting of rheumatoid arthritis, chronic inflammatory bowel disease, chronic inflammatory pelvic disease, multiple sclerosis, asthma, osteoarthritis, psoriasis, atherosclerosis, rhinitis, autoimmunity, and organ transplantation.

The invention also features methods of treating or preventing a disease or disorder by administering to a patient in need of such treatment a substance that modulates the activity of a kinase polypeptide selected from the group consisting of ZC1, ZC2, ZC3, and ZC4. Preferably the disease is selected from the group consisting of immune-related diseases and disorders, cardiovascular disease, and cancer. Most preferably, the immune-related diseases and disorders are selected from the group consisting of rheumatoid arthritis, chronic inflammatory bowel disease, chronic inflammatory pelvic disease, multiple sclerosis, asthma, osteoarthritis, psoriasis, atherosclerosis, rhinitis, autoimmunity, and organ transplantation.

Substances useful for treatment of kinase-related disorders or diseases preferably show positive results in one or more in vitro assays for an activity corresponding to treatment of the disease or disorder in question (Examples of such assays are provided in the references in section VI, below; and in Example 7, herein). Examples of substances that can be screened for favorable activity are provided and referenced in section VI, below. The substances that modulate the activity of the kinases preferably include, but are not limited to, antisense oligonucleotides and inhibitors of protein kinases, as determined by methods and screens referenced in section VI and Example 7, below.

The term “preventing” refers to decreasing the probability that an organism contracts or develops an abnormal condition.

The term “treating” refers to having a therapeutic effect and at least partially alleviating or abrogating an abnormal condition in the organism.

The term “therapeutic effect” refers to the inhibition or activation factors causing or contributing to the abnormal condition. A therapeutic effect relieves to some extent one or more of the symptoms of the abnormal condition. In reference to the treatment of abnormal conditions, a therapeutic effect can refer to one or more of the following: (a) an increase in the proliferation, growth, and/or differentiation of cells; (b) inhibition (i.e., slowing or stopping) of cell death; (c) inhibition of degeneration; (d) relieving to some extent one or more of the symptoms associated with the abnormal condition; and (e) enhancing the function of the affected population of cells. Compounds demonstrating efficacy against abnormal conditions can be identified as described herein.

The term “abnormal condition” refers to a function in the cells or tissues of an organism that deviates from their normal functions in that organism. An abnormal condition can relate to cell proliferation, cell differentiation, or cell survival.

Abnormal cell proliferative conditions include cancers such as fibrotic and mesangial disorders, abnormal angiogenesis and vasculogenesis, wound healing, psoriasis, diabetes mellitus, and inflammation.

Abnormal differentiation conditions include, but are not limited to neurodegenerative disorders, slow wound healing rates, and slow tissue grafting healing rates.

Abnormal cell survival conditions relate to conditions in which programmed cell death (apoptosis) pathways are activated or abrogated. A number of protein kinases are associated with the apoptosis pathways. Aberrations in the function of any one of the protein kinases could lead to cell immortality or premature cell death.

The term “aberration”, in conjunction with the function of a kinase in a signal transduction process, refers to a kinase that is over- or under-expressed in an organism, mutated such that its catalytic activity is lower or higher than wild-type protein kinase activity, mutated such that it can no longer interact with a natural binding partner, is no longer modified by another protein kinase or protein phosphatase, or no longer interacts with a natural binding partner.

The term “administering” relates to a method of incorporating a compound into cells or tissues of an organism. The abnormal condition can be prevented or treated when the cells or tissues of the organism exist within the organism or outside of the organism. Cells existing outside the organism can be maintained or grown in cell culture dishes. For cells harbored within the organism, many techniques exist in the art to administer compounds, including (but not limited to) oral, parenteral, dermal, injection, and aerosol applications. For cells outside of the organism, multiple techniques exist in the art to administer the compounds, including (but not limited to) cell microinjection techniques, transformation techniques, and carrier techniques.

The abnormal condition can also be prevented or treated by administering a compound to a group of cells having an aberration in a signal transduction pathway to an organism. The effect of administering a compound on organism function can then be monitored. The organism is preferably a mouse, rat, rabbit, guinea pig, or goat, more preferably a monkey or ape, and most preferably a human.

In an eleventh aspect, the invention features methods for detection of a kinase polypeptide in a sample as a diagnostic tool for diseases or disorders, wherein the method comprises the steps of: (a) contacting the sample with a nucleic acid probe which hybridizes under hybridization assay conditions to a nucleic acid target region of a kinase polypeptide selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5, said probe comprising the nucleic acid sequence encoding the polypeptide, fragments thereof, and the complements of the sequences and fragments; and (b) detecting the presence or amount of the probe:target region hybrid as an indication of the disease.

In preferred embodiments of the invention, the disease or disorder is selected from the group consisting of rheumatoid arthritis, artherosclerosis, autoimmune disorders, organ transplantation, myocardial infarction, cardiomyopathies, stroke, renal failure, oxidative stress-related neurodegenerative disorders, and cancer. In other preferred embodiments, the kinase polypeptide is selected from the group consisting of PAK4 and PAK5, or the polypeptide is selected from the group consisting of ZC1, ZC2, ZC3, and ZC4, and the disease is cancer.

The kinase “target region” is the nucleotide base sequence set forth in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104, or SEQ ID NO:106, or the corresponding full-length sequences, a functional derivative thereof, or a fragment thereof to which the nucleic acid probe will specifically hybridize. Specific hybridization indicates that in the presence of other nucleic acids the probe only hybridizes detectably with the kinase of the invention's target region. Putative target regions can be identified by methods well known in the art consisting of alignment and comparison of the most closely related sequences in the database.

In preferred embodiments the nucleic acid probe hybridizes to a kinase target region encoding at least 6, 12, 75, 90, 105, 120, 150, 200, 250, 300 or 350 contiguous amino acids of the sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or the corresponding full-length amino acid sequence, or a functional derivative thereof. Hybridization conditions should be such that hybridization occurs only with the kinase genes in the presence of other nucleic acid molecules. Under stringent hybridization conditions only highly complementary nucleic acid sequences hybridize. Preferably, such conditions prevent hybridization of nucleic acids having more than 1 or 2 mismatches out of 20 contiguous nucleotides. Such conditions are defined supra.

The diseases for which detection of kinase genes in a sample could be diagnostic include diseases in which kinase nucleic acid (DNA and/or RNA) is amplified in comparison to normal cells. By “amplification” is meant increased numbers of kinase DNA or RNA in a cell compared with normal cells. In normal cells, kinases are typically found as single copy genes. In selected diseases, the chromosomal location of the kinase genes may be amplified, resulting in multiple copies of the gene, or amplification. Gene amplification can lead to amplification of kinase RNA, or kinase RNA can be amplified in the absence of kinase DNA amplification.

“Amplification” as it refers to RNA can be the detectable presence of kinase RNA in cells, since in some normal cells there is no basal expression of kinase RNA. In other normal cells, a basal level of expression of kinase exists, therefore in these cases amplification is the detection of at least 1-2-fold, and preferably more, kinase RNA, compared to the basal level.

The diseases that could be diagnosed by detection of kinase nucleic acid in a sample preferably include cancers. The test samples suitable for nucleic acid probing methods of the present invention include, for example, cells or nucleic acid extracts of cells, or biological fluids. The samples used in the above-described methods will vary based on the assay format, the detection method and the nature of the tissues, cells or extracts to be assayed. Methods for preparing nucleic acid extracts of cells are well known in the art and can be readily adapted in order to obtain a sample that is compatible with the method utilized.

In a final aspect, the invention features a method for detection of a kinase polypeptide in a sample as a diagnostic tool for a disease or disorder, wherein the method comprises: (a) comparing a nucleic acid target region encoding the kinase polypeptide in a sample, where the kinase polypeptide is selected from the group consisting of STLK2, STLK3, STLK4, STLK5, STLK6, STLK7, ZC1, ZC2, ZC3, ZC4, KHS2, SULU1, SULU3, GEK2, PAK4, and PAK5, or one or more fragments thereof, with a control nucleic acid target region encoding the kinase polypeptide, or one or more fragments thereof; and (b) detecting differences in sequence or amount between the target region and the control target region, as an indication of the disease or disorder. Preferably, the disease or disorder is selected from the group consisting of immune-related diseases and disorders, organ transplantation, myocardial infarction, cardiovascular disease, stroke, renal failure, oxidative stress-related neurodegenerative disorders, and cancer. Immune-related diseases and disorders include, but are not limited to, those discussed previously.

The term “comparing” as used herein refers to identifying discrepancies between the nucleic acid target region isolated from a sample, and the control nucleic acid target region. The discrepancies can be in the nucleotide sequences, e.g. insertions, deletions, or point mutations, or in the amount of a given nucleotide sequence. Methods to determine these discrepancies in sequences are well-known to one of ordinary skill in the art. The “control” nucleic acid target region refers to the sequence or amount of the sequence found in normal cells, e.g. cells that are not diseased as discussed previously.

The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. For example, in some instances the nucleotide sequence of the ZC4 kinase polypeptide may not be part of a preferred embodiment.

The summary of the invention described above is not limiting and other features and advantages of the invention will be apparent from the following detailed description of the invention, and from the claims.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A, 1B and 1C show a multiple sequence alignment of the amino acid sequences (SEQ ID NOS 84-85, 5-7, respectively, in order of appearance) of the STE2O-STE2O family kinases.

FIGS. 2A and 2B show a multiple sequence alignment of the amino acid sequences (SEQ ID NOS 84, 86-87 & 8, respectively, in order of appearance) of the STE2O-STLK5 family kinases.

FIGS. 3A, 3B, 3C, 3D, 3E, 3F and 3G show a multiple sequence alignment of the amino acid sequences (SEQ ID NOS 88-89, 13-16, respectively, in order of appearance) of STE20-ZC family kinases.

FIGS. 4A, 4B and 4C show a pairwise sequence (SEQ ID NOS 91 & 18, respectively, in order of appearance) alignment of STE2O-KHS family kinases.

FIGS. 5A, 5B, 5C and 5D show a multiple sequence alignment of the amino acid sequences (SEQ ID NOS 90, 22, 24 & 151 respectively, in order of appearance) of STE20-SULU family kinases.

FIGS. 6A, 6B and 6C show a pairwise sequence (SEQ ID NOS 92 & 26, respectively, in order of appearance) alignment of STE2O-GEK family kinases

FIGS. 7A, 7B and 7C show a multiple sequence alignment of the amino acid sequences (SEQ ID NOS 93-95, 29-30 respectively, in order of appearance) of STE20-PAK family kinases.

FIGS. 8A, 8B, 8C, 8D, 8E, 8F and 8G show the amino acid sequences of human STLK2(SEQ ID NO:5), human STLK3(SEQ ID NO:6), human STLK4(SEQ ID NO:7), human STLK5(SEQ ID NO:8), human ZC1(SEQ ID NO:13), human ZC2(SEQ ID NO:14), human ZC3(SEQ ID NO:15), human ZC4(SEQ ID NO:16), human KJIS2(SEQ ID NO:18), human SULU1(SEQ ID NO:22), human SULU3(SEQ ID NO:23), murine SULU3(SEQ ID NO:24), human GEK2(SEQ ID NO:26), human PAK4(SEQ ID NO:29), and human PAK5(SEQ ID NO₃₀).

FIGS. 9A, 9B, 9C, 9D, 9E, 9F, 9G, 9H, 9I, 9J, 9K, 9L, 9M, 9N, 9O, 9P, 9Q, 9R, 9S, 9T, 9U and 9V show the nucleic acid sequences of human STLK2(SEQ ID NO:1), human STLK3(SEQ ID NO:2), human STLK4(SEQ ID NO:3), human STLK5(SEQ ID NO:4), human ZC1(SEQ ID NO:9), human ZC2(SEQ ID NO:1O), human ZC3(SEQ ID NO:11), human ZC4(SEQ ID NO:12), human KHS2(SEQ ID NO:17), human SULU1(SEQ ID NO:19), human SULU3(SEQ ID NO:20), murine SULU3(SEQ ID NO:21), human GEK2(SEQ ID NO:25), human PAK4(SEQ ID NO:27), and human PAK5(SEQ ID NO:28).

FIGS. 10A, 10B and 10C show the full-length amino acid sequences of human STLK5 (SEQ ID NO: 97), human PAK5 (SEQ ID NO:1O3), and human ZC4 (SEQ ID NO:105), as well as the partial amino acid sequences of human full-length STLK6 (SEQ ID NO: 99) and human STLK7 (SEQ ID NO: 101) and human GEK2 (SEQ ID NO: 107).

FIGS. 11A, 11B, 11C, 11D, 11E, 11F, 11G and 11H show the full-length nucleic acid sequences of human STLK5 (SEQ ID NO:96), human PAK5 (SEQ ID NO:102), and human ZC4 (SEQ ID NO:104), as well as the partial nucleic acid sequences of human STLK6 (SEQ ID NO: 98) and human STLK7 (SEQ ID NO: 100) and human GEK2 (SEQ ID NO: 106).

FIGS. 12A and 12B show a multiple sequence alignment among human SPAK (SEQ ID NO: 153), human STLK6 (SEQ ID NO: 99), human STLK7 (SEQ ID NO: 101) and full-length human STLK5 (SEQ ID NO: 152).

FIGS. 13A, 13B and 13C show a multiple sequence alignment among human PAKi (SEQ ID NO: 93), human PAK4 (SEQ ID NO: 29) and human PAK5 (SEQ ID NO: 103).

FIGS. 14A, 14B and 14C show a pair-wise sequence alignment between human ZC1 (SEQ ID NO: 15) and human ZC4 (SEQ ID NO: 105).

FIGS. 15A, 15B and 15C show a pair-wise sequence alignment between LOK1 (SEQ ID NO: 154) and full-length GEK2 (SEQ ID NO: 155).

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates in part to kinase polypeptides, nucleic acids encoding such polypeptides, cells containing such nucleic acids, antibodies to such polypeptides, assays utilizing such polypeptides, and methods relating to all of the foregoing. The present invention is based upon the isolation and characterization of new kinase polypeptides. The polypeptides and nucleic acids may be produced using well-known and standard synthesis techniques when given the sequences presented herein.

The recent elucidation of the DNA sequence of Saccharomyces cerevesiae has provided the first complete example of the genetic information contained in a simple eukaryotic organism. Analysis of this yeast genome revealed that it contains at least 113 protein kinases. These kinases were further subdivided into several structurally related groups. One of these newly defined groups was termed the STE20-family to represent its founding member STE20, which is a protein kinase involved in the yeast pheromone response pathway that initiates a protein kinase cascade in response to a G-protein mediated signal. S. cerevesiae has two additional members of this family, CLA4, and YOL113W (HRA655).

Several mammalian homologues have recently been identified that belong to the STE20-family, including SOK-1 (human STE20), GC-kinase, KHS, HPK1, NIK, SLK, GEK, PAK1, PAK65, MST1, and CDC7. Furthermore, the Drosophila and the C. elegans genome efforts have identified additional protein kinases which belong to the STE20-family, yet have structurally unique extracatalytic domains, including ZC504.4 and SULU kinases from C. elegans, and NINAC of Drosophila.

STE20-related protein kinases have been implicated as regulating a variety of cellular responses, including response to growth factors or cytokines, oxidative-, UV-, or irradiation-related stress pathways, inflammatory signals (i.e., TNFα), apoptotic stimuli (i.e., Fas), T and B cell costimulation, the control of cytoskeletal architecture, and cellular transformation. Typically, the STE20-related kinases serve as upstream regulators of MAPK cascades. Examples include: HPK1, a protein-serine/threonine kinase (STK) that possesses a STE20-like kinase domain that activates a protein kinase pathway leading to the stress-activated protein kinase SAPK/JNK; PAK1, an STK with an upstream CDC42-binding domain that interacts with Rac and plays a role in cellular transformation through the Ras-MAPK pathway; and murine NIK, which interacts with upstream receptor tyrosine kinases and connects with downstream STE11-family kinases.

The STE20-kinases possess a variety of non-catalytic domains that are believed to interact with upstream regulators. Examples include proline-rich domains for interaction with SH3-containing proteins, or specific domains for interaction with Rac, Rho, and Rab small G-proteins. These interactions may provide a mechanism for cross-talk between distinct biochemical pathways in response to external stimuli such as the activation of a variety of cell surface receptors, including tyrosine kinases, cytokine receptors, TNF receptor, Fas, T cell receptors, CD28, or CD40.

I. The Nucleic Acids of the Invention

Included within the scope of this invention are the functional equivalents of the herein-described isolated nucleic acid molecules. The degeneracy of the genetic code permits substitution of certain codons by other codons that specify the same amino acid and hence would give rise to the same protein. The nucleic acid sequence can vary substantially since, with the exception of methionine and tryptophan, the known amino acids can be coded for by more than one codon. Thus, portions or all of the kinase genes of the invention could be synthesized to give a nucleic acid sequence significantly different from that shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104, and SEQ ID NO:106. The encoded amino acid sequence thereof would, however, be preserved.

In addition, the nucleic acid sequence may comprise a nucleotide sequence which results from the addition, deletion or substitution of at least one nucleotide to the 5′-end and/or the 3′-end of the nucleic acid formula shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:27, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104, or SEQ ID NO:106, or a derivative thereof. Any nucleotide or polynucleotide may be used in this regard, provided that its addition, deletion or substitution does not alter the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, which is encoded by the nucleotide sequence. For example, the present invention is intended to include any nucleic acid sequence resulting from the addition of ATG as an initiation codon at the 5′-end of the inventive nucleic acid sequence or its derivative, or from the addition of TTA, TAG or TGA as a termination codon at the 3′-end of the inventive nucleotide sequence or its derivative. Moreover, the nucleic acid molecule of the present invention may, as necessary, have restriction endonuclease recognition sites added to its 5′-end and/or 3′-end.

Such functional alterations of a given nucleic acid sequence afford an opportunity to promote secretion and/or processing of heterologous proteins encoded by foreign nucleic acid sequences fused thereto. All variations of the nucleotide sequence of the kinase genes of the invention and fragments thereof permitted by the genetic code are, therefore, included in this invention.

Further, it is possible to delete codons or to substitute one or more codons with codons other than degenerate codons to produce a structurally modified polypeptide, but one which has substantially the same utility or activity as the polypeptide produced by the unmodified nucleic acid molecule. As recognized in the art, the two polypeptides are functionally equivalent, as are the two nucleic acid molecules that give rise to their production, even though the differences between the nucleic acid molecules are not related to the degeneracy of the genetic code.

Mammalian STLK2

The full-length human STLK2 cDNA (SEQ ID NO:1) is 3268 bp long and consists of a 1248 bp open reading frame (ORF) flanked by a 181 bp 5′ untranslated region (UTR; 1-181) and a 1784 bp 3′ UTR (1433-3216) that is followed by a 52 nucleotide polyadenylated region. A polyadenylation signal (AATAAA) is found at positions (3193-3198). The sequence flanking the first ATG conforms to the Kozak consensus (Kozak, M., Nucleic Acids Res. 15, 8125-8148 (1987)) for an initiating methionine, and is believed to be the translational start site for STLK2. Furthermore, human STLK2 and the related SOK-1 and MST3 proteins conserve the amino acid sequence immediately following this presumed initiating methionine.

Several EST fragments span the complete STLK2 sequence with AA191319 at the 5′ end and W16504 at the 3′ end.

Mammalian STLK3

The partial human STLK3 cDNA (SEQ ID NO:2) is 3030 bp long and consists of a 1548 bp ORF flanked by a 1476 bp 3′ UTR (1550-3025) and a 5 nucleotide polyadenylated region. A potential polyadenylation signal (AATAAA) begins at position 3004. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine.

Multiple EST fragments span the complete STLK3 sequence with AA278967 at the 5′ end and AA628477 and others at the 3′ end.

Mammalian STLK4

The partial human STLK4 cDNA (SEQ ID NO:3) is 3857 bp long and consists of a 1242 bp ORF flanked by a 2596 bp 3′ UTR (1244-3839) and an 18 nucleotide polyadenylated region. A potential polyadenylation signal (AATAAA) is found at positions 2181-3822. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. A near full-length murine STLK4 cDNA is represented in the 1773 bp EST AA117438. It extends an additional 21 nucleotides 5′ of the human STLK4 consensus, but since its coding region is open throughout the 5′ extent of the sequence, this is also apparently a partial cDNA clone lacking the N-terminal start methionine.

Several EST fragments span the complete STLK3 sequence with AA297759 at the 5′ end and AA100484 and others at the 3′ end.

Mammalian STLK5

The full-length human STLK5 cDNA (SEQ ID NO:96) is 2110 bp long and consists of a 1119 bp ORF flanked by a 229 bp 5′ UTR and a 762 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (supra) for an initiating methionine, and is believed to be the translational start site for STLK5. Several EST fragments span the complete STLK5 sequence with AA297059 and F07734 at the 5′ end, and R46686 and F03423 and others at the 3′ end.

Mammalian STLK6

The full-length human STLK6 cDNA (SEQ ID NO:98) is 2,001 bp long and consists of a 1,254 bp ORF flanked by a 75 bp 5′ UTR and a 673 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (supra) for an initiating methionine, and is believed to be the translational start site for STLK6.

Mammalian STLK7

The partial human STLK7 cDNA (SEQ ID NO:100) is 311 bp long and consists of a 309 bp ORF. Since the coding region is open throughout both the 5′ and 3′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine and C-terminal stop codon.

Mammalian ZC1

The full-length human ZC1 cDNA (SEQ ID NO:9) is 3798 bp long and consists of a 3717 bp ORF (7-3723) flanked by a 6 bp 5′ UTR and a 75 bp (3724-3798) 3′ UTR. No polyadenylation signal (AATAAA) or polyadenylated region are present in the 3′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human ZC1.

Multiple EST fragments (W81656) match the 3′ end of the human ZC1 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian ZC2

The partial human ZC2 cDNA (SEQ ID NO:10) is 4055 bp long and consists of a 3891 bp ORF (1-3891) and a 164 bp (3892-4055) 3′ UTR. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. No polyadenylation signal (AATAAA) or polyadenylated region are present in the 3′UTR.

Multiple EST fragments (R51245) match the 3′ end of the human ZC2 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian ZC3

The partial human ZC3 cDNA (SEQ ID NO:11) is 4133 bp long and consists of a 3978 bp ORF (1-3978) and a 152 bp (3979-4133) 3′UTR region. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. No polyadenylation signal (AATAAA) or polyadenylated region are present in the 3′UTR.

Multiple EST fragments (R54563) match the 3′end of the human ZC3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian ZC4

The full-length human ZC4 cDNA (SEQ ID NO:104) is 3,684 bp long and was originally assembled from X chromosome genomic DNA sequence.

Multiple EST fragments (R98571) match the 3′end of the human ZC4 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end. ZC4 gene is also contained within the human genomic clone Z83850.

Mammalian KHS2

The full-length human KHS2 cDNA (SEQ ID NO:17) is 4023 bp long and consists of a 2682 bp ORF (6-2687) flanked by a 5 bp (1-5) 5′UTR and a 1336 bp (2688-4023) 3′ UTR. A potential polyadenylation signal (AATAAA) is found at positions 4008-4013. No polyadenylated region is present in the 3′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human KHS2.

Multiple EST fragments match the 5′end (AA446022) as well as the 3′ end (R37625) of the human KHS2 gene.

Mammalian SULU1

The full-length human SULU1 cDNA (SEQ ID NO:19) is 4177 bp long and consists of a 2694 bp ORF (415-3108) flanked by a 414 bp (1-414) 5′UTR and a 1069 bp (3109-4177) 3′ UTR followed by a 19 nucleotide polydenylated region. A potential polyadenylation signal (AATAAA) is found at positions 4164-4169. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human SULU1.

Multiple EST fragments match the 5′end (N27153) as well as the 3′ end (R90908) of the human SULU1 gene.

Mammalian (Murine) SULU3

The partial murine SULU3 cDNA (SEQ ID NO:21) is 2249 bp long and consists of a 2244 bp ORF (6-2249) flanked by a 5 bp (1-5) 5′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for murine SULU3. The 3′ end of the murine SULU3 cDNA shares 90% DNA sequence identity over 1620 nucleotides with human SULU3, suggesting that these two genes are functional orthologues.

One EST fragment (AA446022) matches the 3′ end of the partial murine SULU3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian (Human) SULU3

The partial human SULU3 cDNA (SEQ ID NO:20) is 3824 bp long and consists of a 2358 bp ORF (2-2359) flanked by a 1465 bp (2360-3824) 3′UTR followed by a 19 nucleotide polydenylated region. A potential polyadenylation signal (AATAAA) is found at positions 2602-2607. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. The 5′ end of the human SULU3 cDNA shares 90% DNA sequence identity over 1620 nucleotides with murine SULU3, suggesting that these two genes are functional orthologues.

Multiple EST fragments (R02283) match the 3′end of the human SULU3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian GEK2

The full-length human GEK2 cDNA (SEQ ID NO:106) is 2962 bp long and consists of a 2737 bp ORF (59-2795) flanked by a 58 bp (1-58) 5′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human GEK2.

Multiple EST fragments (AA465671) match the 5′end, but at the time of filing, the inventors believe that only one (AA380492) matches the 3′ end of the human GEK2 gene.

Mammalian PAK4

The full-length human PAK4 cDNA (SEQ ID NO:27) is 3604 bp long and consists of a 2043 bp ORF (143-2185) flanked by a 142 bp (1-142) 5′UTR and a 1419 3′ UTR followed by a 22 nucleotide polydenylated region. A potential polyadenylation signal (AATTAAA) is found at positions 3582-3588. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human PAK4.

Multiple EST fragments (AA535791) match the 3′end of the human PAK4 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

Mammalian PAK5

The full-length human PAK5 cDNA (SEQ ID NO:102) is 2806 bp long and consists of a 1773 bp ORF flanked by a 201 bp 5′ UTR and a 833 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (supra) for an initiating methionine, and is believed to be the translational start site for PAK5.

Multiple EST fragments (AA442867) match the 3′end of the human PAK5 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

II. Nucleic Acid Probes, Methods, and Kits for Detection of STE20-Related Kinases.

A nucleic acid probe of the present invention may be used to probe an appropriate chromosomal or cDNA library by usual hybridization methods to obtain other nucleic acid molecules of the present invention. A chromosomal DNA or cDNA library may be prepared from appropriate cells according to recognized methods in the art (cf. “Molecular Cloning: A Laboratory Manual”, second edition, Cold Spring Harbor Laboratory, Sambrook, Fritsch, & Maniatis, eds., 1989).

In the alternative, chemical synthesis can be carried out in order to obtain nucleic acid probes having nucleotide sequences which correspond to N-terminal and C-terminal portions of the amino acid sequence of the polypeptide of interest. The synthesized nucleic acid probes may be used as primers in a polymerase chain reaction (PCR) carried out in accordance with recognized PCR techniques, essentially according to PCR Protocols, “A Guide to Methods and Applications”, Academic Press, Michael, et al., eds., 1990, utilizing the appropriate chromosomal or cDNA library to obtain the fragment of the present invention.

One skilled in the art can readily design such probes based on the sequence disclosed herein using methods of computer alignment and sequence analysis known in the art (“Molecular Cloning: A Laboratory Manual”, 1989, supra). The hybridization probes of the present invention can be labeled by standard labeling techniques such as with a radiolabel, enzyme label, fluorescent label, biotin-avidin label, chemiluminescence, and the like. After hybridization, the probes may be visualized using known methods.

The nucleic acid probes of the present invention include RNA, as well as DNA probes, such probes being generated using techniques known in the art. The nucleic acid probe may be immobilized on a solid support. Examples of such solid supports include, but are not limited to, plastics such as polycarbonate, complex carbohydrates such as agarose and sepharose, and acrylic resins, such as polyacrylamide and latex beads. Techniques for coupling nucleic acid probes to such solid supports are well known in the art.

The test samples suitable for nucleic acid probing methods of the present invention include, for example, cells or nucleic acid extracts of cells, or biological fluids. The samples used in the above-described methods will vary based on the assay format, the detection method and the nature of the tissues, cells or extracts to be assayed. Methods for preparing nucleic acid extracts of cells are well known in the art and can be readily adapted in order to obtain a sample which is compatible with the method utilized.

One method of detecting the presence of nucleic acids of the invention in a sample comprises (a) contacting said sample with the above-described nucleic acid probe under conditions such that hybridization occurs, and (b) detecting the presence of said probe bound to said nucleic acid molecule. One skilled in the art would select the nucleic acid probe according to techniques known in the art as described above. Samples to be tested include but should not be limited to RNA samples of human tissue.

A kit for detecting the presence of nucleic acids of the invention in a sample comprises at least one container means having disposed therein the above-described nucleic acid probe. The kit may further comprise other containers comprising one or more of the following: wash reagents and reagents capable of detecting the presence of bound nucleic acid probe. Examples of detection reagents include, but are not limited to radiolabelled probes, enzymatic labeled probes (horseradish peroxidase, alkaline phosphatase), and affinity labeled probes (biotin, avidin, or steptavidin).

In detail, a compartmentalized kit includes any kit in which reagents are contained in separate containers. Such containers include small glass containers, plastic containers or strips of plastic or paper. Such containers allow the efficient transfer of reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another. Such containers will include a container which will accept the test sample, a container which contains the probe or primers used in the assay, containers which contain wash reagents (such as phosphate buffered saline, Tris-buffers, and the like), and containers which contain the reagents used to detect the hybridized probe, bound antibody, amplified product, or the like. One skilled in the art will readily recognize that the nucleic acid probes described in the present invention can readily be incorporated into one of the established kit formats which are well known in the art.

III. DNA Constructs Comprising a STE20-Related Nucleic Acid Molecule and Cells Containing These Constructs.

The present invention also relates to a recombinant DNA molecule comprising, 5′ to 3′, a promoter effective to initiate transcription in a host cell and the above-described nucleic acid molecules. In addition, the present invention relates to a recombinant DNA molecule comprising a vector and an above-described nucleic acid molecule. The present invention also relates to a nucleic acid molecule comprising a transcriptional region functional in a cell, a sequence complementary to an RNA sequence encoding an amino acid sequence corresponding to the above-described polypeptide, and a transcriptional termination region functional in said cell. The above-described molecules may be isolated and/or purified DNA molecules.

The present invention also relates to a cell or organism that contains an above-described nucleic acid molecule and thereby is capable of expressing a polypeptide. The polypeptide may be purified from cells which have been altered to express the polypeptide. A cell is said to be “altered to express a desired polypeptide” when the cell, through genetic manipulation, is made to produce a protein which it normally does not produce or which the cell normally produces at lower levels. One skilled in the art can readily adapt procedures for introducing and expressing either genomic, cDNA, or synthetic sequences into either eukaryotic or prokaryotic cells.

A nucleic acid molecule, such as DNA, is said to be “capable of expressing” a polypeptide if it contains nucleotide sequences which contain transcriptional and translational regulatory information and such sequences are “operably linked” to nucleotide sequences which encode the polypeptide. An operable linkage is a linkage in which the regulatory DNA sequences and the DNA sequence sought to be expressed are connected in such a way as to permit gene sequence expression. The precise nature of the regulatory regions needed for gene sequence expression may vary from organism to organism, but shall in general include a promoter region which, in prokaryotes, contains both the promoter (which directs the initiation of RNA transcription) as well as the DNA sequences which, when transcribed into RNA, will signal synthesis initiation. Such regions will normally include those 5′-non-coding sequences involved with initiation of transcription and translation, such as the TATA box, capping sequence, CAAT sequence, and the like.

If desired, the non-coding region 3′ to the sequence encoding a kinase of the invention may be obtained by the above-described methods. This region may be retained for its transcriptional termination regulatory sequences, such as termination and polyadenylation. Thus, by retaining the 3′-region naturally contiguous to the DNA sequence encoding a kinase of the invention, the transcriptional termination signals may be provided. Where the transcriptional termination signals are not satisfactorily functional in the expression host cell, then a 3′ region functional in the host cell may be substituted.

Two DNA sequences (such as a promoter region sequence and a sequence encoding a kinase of the invention) are said to be operably linked if the nature of the linkage between the two DNA sequences does not (1) result in the introduction of a frame-shift mutation, (2) interfere with the ability of the promoter region sequence to direct the transcription of a gene sequence encoding a kinase of the invention, or (3) interfere with the ability of the gene sequence of a kinase of the invention to be transcribed by the promoter region sequence. Thus, a promoter region would be operably linked to a DNA sequence if the promoter were capable of effecting transcription of that DNA sequence. Thus, to express a gene encoding a kinase of the invention, transcriptional and translational signals recognized by an appropriate host are necessary.

The present invention encompasses the expression of a gene encoding a kinase of the invention (or a functional derivative thereof) in either prokaryotic or eukaryotic cells. Prokaryotic hosts are, generally, very efficient and convenient for the production of recombinant proteins and are, therefore, one type of preferred expression system for kinases of the invention. Prokaryotes most frequently are represented by various strains of E. coli. However, other microbial strains may also be used, including other bacterial strains.

In prokaryotic systems, plasmid vectors that contain replication sites and control sequences derived from a species compatible with the host may be used. Examples of suitable plasmid vectors may include pBR322, pUC118, pUC119 and the like; suitable phage or bacteriophage vectors may include γgt10, γgt11 and the like; and suitable virus vectors may include pMAM-neo, PKRC and the like. Preferably, the selected vector of the present invention has the capacity to replicate in the selected host cell.

Recognized prokaryotic hosts include bacteria such as E. coli, Bacillus, Streptomyces, Pseudomonas, Salmonella, Serratia, and the like. However, under such conditions, the polypeptide will not be glycosylated. The prokaryotic host must be compatible with the replicon and control sequences in the expression plasmid.

To express a kinase of the invention (or a functional derivative thereof) in a prokaryotic cell, it is necessary to operably link the sequence encoding the kinase of the invention to a functional prokaryotic promoter. Such promoters may be either constitutive or, more preferably, regulatable (i.e., inducible or derepressible). Examples of constitutive promoters include the int promoter of bacteriophage λ, the bla promoter of the β-lactamase gene sequence of pBR322, and the cat promoter of the chloramphenicol acetyl transferase gene sequence of pPR325, and the like. Examples of inducible prokaryotic promoters include the major right and left promoters of bacteriophage λ (P_(L) and P_(R)), the trp, recA, λacZ, λacI, and gal promoters of E. coli, the α-amylase (Ulmanen et al., J. Bacteriol. 162:176-182, 1985) and the ζq-28-specific promoters of B. subtilis (Gilman et al., Gene Sequence 32:11-20, 1984), the promoters of the bacteriophages of Bacillus (Gryczan, In: The Molecular Biology of the Bacilli, Academic Press, Inc., NY, 1982), and Streptomyces promoters (Ward et al., Mol. Gen. Genet. 203:468-478, 1986). Prokaryotic promoters are reviewed by Glick (Ind. Microbiot. 1:277-282, 1987), Cenatiempo (Biochimie 68:505-516, 1986), and Gottesman (Ann. Rev. Genet. 18:415-442, 1984).

Proper expression in a prokaryotic cell also requires the presence of a ribosome-binding site upstream of the gene sequence-encoding sequence. Such ribosome-binding sites are disclosed, for example, by Gold et al. (Ann. Rev. Microbiol. 35:365-404, 1981). The selection of control sequences, expression vectors, transformation methods, and the like, are dependent on the type of host cell used to express the gene. As used herein, “cell”, “cell line”, and “cell culture” may be used interchangeably and all such designations include progeny. Thus, the words “transformants” or “transformed cells” include the primary subject cell and cultures derived therefrom, without regard to the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. However, as defined, mutant progeny have the same functionality as that of the originally transformed cell.

Host cells which may be used in the expression systems of the present invention are not strictly limited, provided that they are suitable for use in the expression of the kinase polypeptide of interest. Suitable hosts may often include eukaryotic cells. Preferred eukaryotic hosts include, for example, yeast, fungi, insect cells, mammalian cells either in vivo, or in tissue culture. Mammalian cells which may be useful as hosts include HeLa cells, cells of fibroblast origin such as VERO or CHO-K1, or cells of lymphoid origin and their derivatives. Preferred mammalian host cells include SP2/0 and J558L, as well as neuroblastoma cell lines such as IMR 332, which may provide better capacities for correct post-translational processing.

In addition, plant cells are also available as hosts, and control sequences compatible with plant cells are available, such as the cauliflower mosaic virus 35S and 19S, and nopaline synthase promoter and polyadenylation signal sequences. Another preferred host is an insect cell, for example the Drosophila larvae. Using insect cells as hosts, the Drosophila alcohol dehydrogenase promoter can be used (Rubin, Science 240:1453-1459, 1988). Alternatively, baculovirus vectors can be engineered to express large amounts of kinases of the invention in insect cells (Jasny, Science 238:1653, 1987; Miller et al., In: Genetic Engineering, Vol. 8, Plenum, Setlow et al., eds., pp. 277-297, 1986).

Any of a series of yeast expression systems can be utilized which incorporate promoter and termination elements from the actively expressed sequences coding for glycolytic enzymes that are produced in large quantities when yeast are grown in mediums rich in glucose. Known glycolytic gene sequences can also provide very efficient transcriptional control signals. Yeast provides substantial advantages in that it can also carry out post-translational modifications. A number of recombinant DNA strategies exist utilizing strong promoter sequences and high copy number plasmids which can be utilized for production of the desired proteins in yeast. Yeast recognizes leader sequences on cloned mammalian genes and secretes peptides bearing leader sequences (i.e., pre-peptides). Several possible vector systems are available for the expression of kinases of the invention in a mammalian host.

A wide variety of transcriptional and translational regulatory sequences may be employed, depending upon the nature of the host. The transcriptional and translational regulatory signals may be derived from viral sources, such as adenovirus, bovine papilloma virus, cytomegalovirus, simian virus, or the like, where the regulatory signals are associated with a particular gene sequence which has a high level of expression. Alternatively, promoters from mammalian expression products, such as actin, collagen, myosin, and the like, may be employed. Transcriptional initiation regulatory signals may be selected which allow for repression or activation, so that expression of the gene sequences can be modulated. Of interest are regulatory signals which are temperature-sensitive so that by varying the temperature, expression can be repressed or initiated, or are subject to chemical (such as metabolite) regulation.

Expression of kinases of the invention in eukaryotic hosts requires the use of eukaryotic regulatory regions. Such regions will, in general, include a promoter region sufficient to direct the initiation of RNA synthesis. Preferred eukaryotic promoters include, for example, the promoter of the mouse metallothionein I gene sequence (Hamer et al., J. Mol. Appl. Gen. 1:273-288, 1982); the TK promoter of Herpes virus (McKnight, Cell 31:355-365, 1982); the SV40 early promoter (Benoist et al., Nature (London) 290:304-31, 1981); and the yeast gal4 gene sequence promoter (Johnston et al., Proc. Natl. Acad. Sci. (USA) 79:6971-6975, 1982; Silver et al., Proc. Natl. Acad. Sci. (USA) 81:5951-5955, 1984).

Translation of eukaryotic mRNA is initiated at the codon which encodes the first methionine. For this reason, it is preferable to ensure that the linkage between a eukaryotic promoter and a DNA sequence which encodes a kinase of the invention (or a functional derivative thereof) does not contain any intervening codons which are capable of encoding a methionine (i.e., AUG). The presence of such codons results either in the formation of a fusion protein (if the AUG codon is in the same reading frame as the kinase of the invention coding sequence) or a frame-shift mutation (if the AUG codon is not in the same reading frame as the kinase of the invention coding sequence).

A nucleic acid molecule encoding a kinase of the invention and an operably linked promoter may be introduced into a recipient prokaryotic or eukaryotic cell either as a nonreplicating DNA or RNA molecule, which may either be a linear molecule or, more preferably, a closed covalent circular molecule. Since such molecules are incapable of autonomous replication, the expression of the gene may occur through the transient expression of the introduced sequence. Alternatively, permanent expression may occur through the integration of the introduced DNA sequence into the host chromosome.

A vector may be employed which is capable of integrating the desired gene sequences into the host cell chromosome. Cells which have stably integrated the introduced DNA into their chromosomes can be selected by also introducing one or more markers which allow for selection of host cells which contain the expression vector. The marker may provide for prototrophy to an auxotrophic host, biocide resistance, e.g., antibiotics, or heavy metals, such as copper, or the like. The selectable marker gene sequence can either be directly linked to the DNA gene sequences to be expressed, or introduced into the same cell by co-transfection. Additional elements may also be needed for optimal synthesis of mRNA. These elements may include splice signals, as well as transcription promoters, enhancers, and termination signals. cDNA expression vectors incorporating such elements include those described by Okayama (Mol. Cell. Biol. 3:280-, 1983).

The introduced nucleic acid molecule can be incorporated into a plasmid or viral vector capable of autonomous replication in the recipient host. Any of a wide variety of vectors may be employed for this purpose. Factors of importance in selecting a particular plasmid or viral vector include: the ease with which recipient cells that contain the vector may be recognized and selected from those recipient cells which do not contain the vector; the number of copies of the vector which are desired in a particular host; and whether it is desirable to be able to “shuttle” the vector between host cells of different species.

Preferred prokaryotic vectors include plasmids such as those capable of replication in E. coli (such as, for example, pBR322, ColEl, pSC101, pACYC 184, πVX; “Molecular Cloning: A Laboratory Manual”, 1989, supra). Bacillus plasmids include pCl94, pC221, pT127, and the like (Gryczan, In: The Molecular Biology of the Bacilli, Academic Press, NY, pp. 307-329, 1982). Suitable Streptomyces plasmids include p1J101 (Kendall et al., J. Bacteriol. 169:4177-4183, 1987), and streptomyces bacteriophages such as φC31 (Chater et al., In: Sixth International Symposium on Actinomycetales Biology, Akademiai Kaido, Budapest, Hungary, pp. 45-54, 1986). Pseudomonas plasmids are reviewed by John et al. (Rev. Infect. Dis. 8:693-704, 1986), and Izaki (Jpn. J. Bacteriol. 33:729-742, 1978).

Preferred eukaryotic plasmids include, for example, BPV, vaccinia, SV40, 2-micron circle, and the like, or their derivatives. Such plasmids are well known in the art (Botstein et al., Miami Wntr. Symp. 19:265-274, 1982; Broach, In: “The Molecular Biology of the Yeast Saccharomyces: Life Cycle and Inheritance”, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., p. 445-470, 1981; Broach, Cell 28:203-204, 1982; Bollon et al., J. Clin. Hematol. Oncol. 10:39-48, 1980; Maniatis, In: Cell Biology: A Comprehensive Treatise, Vol. 3, Gene Sequence Expression, Academic Press, NY, pp. 563-608, 1980).

Once the vector or nucleic acid molecule containing the construct(s) has been prepared for expression, the DNA construct(s) may be introduced into an appropriate host cell by any of a variety of suitable means, i.e., transformation, transfection, conjugation, protoplast fusion, electroporation, particle gun technology, calcium phosphate-precipitation, direct microinjection, and the like. After the introduction of the vector, recipient cells are grown in a selective medium, which selects for the growth of vector-containing cells. Expression of the cloned gene(s) results in the production of a kinase of the invention, or fragments thereof. This can take place in the transformed cells as such, or following the induction of these cells to differentiate (for example, by administration of bromodeoxyuracil to neuroblastoma cells or the like). A variety of incubation conditions can be used to form the peptide of the present invention. The most preferred conditions are those which mimic physiological conditions.

IV. The Proteins of the Invention

A variety of methodologies known in the art can be utilized to obtain the polypeptides of the present invention. The polypeptides may be purified from tissues or cells that naturally produce the polypeptides. Alternatively, the above-described isolated nucleic acid fragments could be used to express the kinases of the invention in any organism. The samples of the present invention include cells, protein extracts or membrane extracts of cells, or biological fluids. The samples will vary based on the assay format, the detection method, and the nature of the tissues, cells or extracts used as the sample.

Any eukaryotic organism can be used as a source for the polypeptides of the invention, as long as the source organism naturally contains such polypeptides. As used herein, “source organism” refers to the original organism from which the amino acid sequence of the subunit is derived, regardless of the organism the subunit is expressed in and ultimately isolated from.

One skilled in the art can readily follow known methods for isolating proteins in order to obtain the polypeptides free of natural contaminants. These include, but are not limited to: size-exclusion chromatography, HPLC, ion-exchange chromatography, and immuno-affinity chromatography.

Mammalian STLK2

Analysis of the deduced amino acid sequence predicts STLK2 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. STLK2 contains a 21 amino acid N-terminal domain, a 253 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, followed by a 142 amino acid C-terminal domain.

STLK2 is most closely related to human STE20-subfamily kinases, MST3 (GB:AF024636) and SOK-1 (GB:X99325) and a C. elegans kinase yk34b11.5 (GB:U53153) sharing 72.7%, 68.7%, and 69.3% amino acid identity, respectively.

The 21 amino acid N-terminal domain of human STLK2 is 71.4% identical to the N-terminus of MST3 (GB:AF024636). Human STLK2 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 253 amino acid catalytic domain of human STLK2 is most related to human SOK-1 (X99325), MST3 (GB:AF024636), C. elegans yk32b11.5 (GB:U53153), and STLK3 (SEQ ID NO:6) sharing 88.9%, 87.4%, 78.3%, and 49% amino identity respectively, placing it in the STLK-subfamily of STE20-related kinases. The STLK2 kinase domain displayed lesser homology to other STE20-related kinases including: 55.9% to human MST2 (GB:U26424), 49.2% to human GCK (GB:U07349), 49.2% to human KHS1 (GB:U77129), and 44.2% to human HPKl (GB:U66464). The activation loop of human STLK2 catalytic domain is identical to that of human SOK-1 and MST3 including the presence of four potential threonine phosphorylation sites that could serve an autoregulatory role on kinase activity.

The 142 amino acid C-terminal domain of human STLK2 is most related to human SOK-1 (X99325), MST3 (GB:AF024636), and C. elegans yk32b11.5 (GB:U53153), sharing 39.9%, 39.9%, and 33.3% amino acid identity, respectively. This C-terminal domain shares some significant amino acid similarity to the C-terminal domains of the related human STLK3 (SEQ ID NO:6) and STLK4 (SEQ ID NO:7).

The C-terminus of the related human SOK-1 (GB:X99325) kinase has been shown to be inhibitory to the catalytic activity of this kinase (Pombo, C. M., Bonventre, J. V., Molnar, A., Kyriakis, J. and Force, T. EMBO J. 15, 4537-4546 (1996)). Based on the sequence identity between the C-termini of human SOK-1 (GB:X99325) and human STLK2 (39.2%), the C-terminus of human STLK2 may also function as an inhibitory domain for its kinase.

Mammalian STLK3

The 3030 bp human STLK3 nucleotide sequence of the partial cDNA clone encodes a polypeptide of 516 amino acids (SEQ ID NO:6) with a predicted molecular mass of 56,784 daltons. Analysis of the deduced amino acid sequence predicts STLK3 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain, however the cDNA clone lacks an initiating ATG, so the full extent of it N-termius is not known. STLK3 contains a 31 amino acid N-terminal domain, a 277 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, followed by a 181 amino acid C-terminal domain containing a 25 amino acid insert and a 27 amino acid tail relative to the sequence of human STLK2.

STLK3 is most closely related to human STE20-subfamily kinases, STLK4 (SEQ ID. NO:7), MST3 (GB:AF024636), SOK-1 (GB:X99325) and STLK2 (SEQ ID NO:5) sharing 71.1%, 37.6%, 38.1%, and 38.4% amino acid identity respectively.

The 31 amino acid N-terminal domain of human STLK3 lacked any significant amino acid sequence homologies using a Smith-Waterman search of the nonredundant protein database, other than sequence similarity to proline-alanine repeats.

The 277 amino acid catalytic domain of human STLK3 is most related to human STLK4 (SEQ ID NO:7), SOK-1 (GB:X99325), MST3 (GB:AF024636), and STLK2 (SEQ ID NO:5) sharing 88.2%, 49.2%, 49%, and 49% amino acid identity, respectively. It also shares strong homology to other STKs from lower organisms including 51.7% to A. thaliana (GB: AC002343), 43.1% to A. thaliana (GB: Z97336), 42.1% to A. thaliana (GB: U96613), and 43.3% to C. elegans (GB: U53153). The activation loop of the human STLK3 catalytic domain conserves three potential threonine phosphorylation sites with other members of the STLK-subfamily of STE20-related kinases (human STE20, MST3, STLK2, STLK4) that could serve an autoregulatory role on kinase activity.

The 181 amino acid C-terminal domain of human STLK3 shares 55.5% amino acid identity to human STLK4 (SEQ ID NO:7), and is 100% identical to a partial human cDNA DCHT (GB:AF017635). The C-terminal domain of human STLK3 contains a 26 amino acid insert relative to human STE20. A similar (87.5% amino acid identity) 26 amino acid insert is also present in human STLK4.

The 27 amino acid C-terminal tail of human STLK3 shares 77.8% amino acid identity to human STLK4, but is absent from other STLK-family members. This high degree of homology between the C-tail of two STLK-family members suggests they may be involved in an as yet unidentified protein—protein interaction.

The weak sequence homology between the C-termini of human STLK3 and STE20, suggests it may also function as an inhibitory domain for its kinase.

Mammalian STLK4

The 3857 bp human STLK4 nucleotide sequence of the partial cDNA clone encodes a polypeptide of 414 amino acids (SEQ ID NO:7) with a predicted molecular mass of 45,451 daltons. Analysis of the deduced amino acid sequence predicts STLK4 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain, however the cDNA clone lacks an initiating ATG, so the full extent of it N-terminus is not known. The partial STLK4 protein sequence contains a 178 amino acid catalytic domain corresponding to the C-terminal motifs VI-XI of a serine/threonine kinase, followed by a 236 amino acid C-terminal domain containing two inserts of 25 and 41 amino acids each, relative to the sequence of human STLK2.

STLK4 is most closely related to human STE20-subfamily kinases, STLK3 (SEQ ID. NO 6), MST3 (GB:AF024636), STLK2 (SEQ ID NO:5), and SOK-1 (GB:X99325) sharing 71.0%, 46.8%, 43.9%, and 37.7% amino acid identity, respectively.

The 178 amino acid catalytic domain of human STLK4 is most related to human STLK3 (SEQ ID NO. 7), SOK-1 (GB:X99325), MST3 (GB:AF024636), STLK2 (SEQ ID NO:5), and MST1 (GB:U18297), sharing 88.2%, 54.2%, 54.0%, 53.7 and 45.7% amino acid identity, respectively. It also shares strong homology to other STKs from lower organisms including 56.9% to A. thaliana (GB: AC002343), 52.5% to C. elegans (GB: U53153), 46.2% to A. thaliana (GB: Z97336) and 45.7% to A. thaliana (GB: U96613). The activation loop of the human STLK4 catalytic domain conserves three potential threonine phosphorylation sites with other members of the STLK-subfamily of STE20-related kinases (human STE20, MST3, STLK2 and STLK3) that could serve an autoregulatory role on kinase activity.

The 236 amino acid C-terminal domain of human STLK4 shares 58.1% amino acid identity to both human STLK3 (SEQ ID NO:6) and to a partial human cDNA, DCHT (GB:AF017635). The C-terminal domain of human STLK4 contains a 25 amino acid insert relative to human SOK-1 and shares 87.5% amino acid identity to an insert present in human STLK3.

The weak sequence homology between the C-termini of human STLK4 and STE20, suggests it may also function as an inhibitory domain for its kinase.

Mammalian STLK5

The full-length 2110 bp human STLK5 cDNA encodes a polypeptide of 373 amino acids (SEQ ID NO:97) with a predicted molecular mass of 41,700 daltons. Analysis of the deduced amino acid sequence predicts STLK5 to be an intracellular STE20-subfamily kinase, lacking both a signal sequence and transmembrane domain. STLK5 contains a 10 amino acid N-terminal domain, a 311 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, and a 52 amino acid C-terminal domain.

STLK5 is most closely related to the human STE20-subfamily kinases STLK6 (SEQ ID No. 99) and SPAK (AFO99989), sharing 51% and 33% amino acid identity, respectively, over its full extent. It also shares significant homology to database entries from Arabidopsis thaliana (GB:AC002343) and C.elegans (GB:AL023843, GB:AL023843).

The 10 amino acid N-terminal domain of human STLK5 does not reveal any significant homologies to the protein database.

The 311 amino acid catalytic domain of human STLK5 shares 51% and 34% identity to STLK6 and SPAK, respectively. The catalytic domain of STLK5 contains a 45 amino acid insert between kinase subdomains X and XI relative to human STE20. Multiple human EST fragments as well as a murine EST (GB:AA575647) contain this insert providing evidence that this region is an integral part of STLK5.

The 52 amino acid C-terminal tail of human STLK5 shares 41.3% amino acid identity to human SOK-1 (GB:X99325). The weak sequence homology between the C-termini of human STLK5 and STE20, suggests it may also function as an inhibitory domain for its kinase.

Mammalian STLK6

The 2,001 bp human STLK6 nucleotide sequence of the complete cDNA encodes a polypeptide of 418 amino acids (SEQ ID NO:99) with a predicted molecular mass of 47,025 daltons. Analysis of the deduced amino acid sequence predicts STLK6 to be an intracellular STE20-subfamily kinase, lacking both a signal sequence and transmembrane domain. STLK6 contains a 57 amino acid N-terminal domain, a 312 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, followed by a 49 amino acid C-terminal domain.

STLK6 is most closely related to human STE20-subfamily kinases STLK5 (SEQ ID NO:97), STLK7 (SEQ ID NO:101), and SPAK (AF099989), sharing 50%, 35%, and 30% amino acid identity over its full extent. It also shares significant homology to database entries from Arabidopsis thaliana (GB:AC002343) and C.elegans (GB:U53153). The 57 amino acid N-terminal domain of human STLK6 does not reveal any significant homologies in the protein database.

The 312 amino acid catalytic domain of human STLK6 shares 51 and 30% identity to human STLK5 and SPAK, respectively.

The 49 amino acid C-terminal tail of human STLK6 shares low amino acid sequence identity (29%) with STLK5 and SPAK.

Mammalian STLK7

The 311 bp human STLK7 nucleotide sequence of the partial cDNA encodes a polypeptide of 103 amino acids (SEQ ID NO:101). Analysis of the deduced amino acid sequence predicts STLK7 to be an internal fragment of an intracellular STE20-family kinase. This sequence lacks the N- and C-terminal portions of STLK7 and contains only the N-terminal 103 amino acids of the predicted catalytic domain.

Human STLK7 is most closely related to human STE20-subfamily kinases SPAK (AF099989), STLK5 (SEQ ID NO:97), and STLK6 (SEQ ID NO:99), sharing 86%, 38%, and 35% amino acid identity within this region of the kinase domain. It also shares significant homology to database entries from Arabidopsis thaliana (GB:AC002343) and Drosophila melanogaster (GB:AF006640).

Mammalian ZC1

The 3798 bp human ZC1 nucleotide sequence encodes a polypeptide of 1239 amino acids (SEQ ID NO:13) with a predicted molecular mass of 142,140 daltons. Analysis of the deduced amino acid sequence predicts ZC1 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The full-length ZC1 protein contains a 22 amino acid N-terminus, a 267 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 237 amino acid region predicted to form a coiled-coil structure, a 114 amino acid proline-rich region, a 256 amino acid spacer region, followed by a 343 amino acid C-terminal domain containing a potential Rab/Rho-binding region.

ZC1 is most closely related to the human STE20-subfamily kinases ZC2 (SEQ ID NO:14), ZC3 (SEQ ID NO:15), and ZC4 (SEQ ID NO:16), sharing 61.7%, 60.9%, and 43.8% amino acid identity, respectively. ZC1 also shares 45.5% amino acid identity to a C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029). ZC1 exhibits 90.0% amino acid homology to murine NIK (GB:U88984), suggesting it may be the human orthologue of this STK.

The 22 amino acid N-terminal domain of human ZC1 is 58.8% identical to the C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029), and 100% identical to murine NIK (GB: U88984). Human ZC1 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 267 amino acid catalytic domain of human ZC1 is most related to human STE20-subfamily kinases, ZC3 (SEQ ID NO:15), ZC2 (SEQ ID NO:14), KHS2 (SEQ ID NO:18), SOK-1 (GB:X99325), GCK (GB:U07349), and GEK2 (SEQ ID NO:107), and to the C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029) sharing 90.6%, 90.2%, 50.6%, 47.4%, 45.4%, 42.5% and 82.6% amino acid identity, respectively. The ZC1 kinase domain shares 98.1% identity to murine NIK (GB:U88984). ZC1 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3, PAK4 and PAK5.

Immediately C-terminal to the kinase domain of human ZC1 is a 237 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm (Lupas, A. Meth. Enzymol. 266, 513-525 (1996)). This region of ZC1 is most related to human STE20-subfamily kinases, ZC3 (SEQ ID NO:15), ZC2 (SEQ ID NO:14), and GEK2 (SEQ ID NO:107), as well as to human PITSLRE (GB:U04824) sharing 65.5%, 65.4%, 25.3%, and 29.0% amino acid identity, respectively. The ZC1 coiled-coil domain also shares 90.6% amino acid homology to murine NIK. The C. elegans homologue ZC504.4 shares 32.2% sequence identity over this region.

Within the predicted coiled-coil domain of human ZCl, and the related ZC3, is a region predicted to form a leucine zipper (Leu-X6-Leu-X6-Leu-X6-Leu-X20-Leu-X6-Leu) (SEQ ID NO: 149). The fact that this leucine repeat exists within a predicted coiled-coil structure suggests that the leucine zipper may have a high probability of serving as a dimerization interface (Hirst, J. D. et al Protein Engineering 9 657-662 (1996)) mediating a potential inter- or intra-molecular dimerization of human ZC1.

The 114 amino acid proline-rich region of human ZC1 is most related to human STE20-subfamily kinases, ZC2 (SEQ ID NO:14) and ZC3 (SEQ ID NO:15), sharing 35.8%, and 24.9%, respectively. The ZC1 proline-rich domain shares 36.4% amino acid homology to murine NIK (GB:U88984). Three potential “PxxP” (SEQ ID NO: 148) SH3 domain-binding motifs (I, II and III) are found within the proline-rich region of human ZC 1. Motif I is conserved in human ZC1 and C. elegans ZC504.4 (GB:Z50029). Motif II is conserved in ZC1, ZC2, ZC3, ZC4 and C. elegans ZC504.4. Motif III is conserved in ZC1, ZC2, ZC3 and ZC4. Motifs II and III of murine NIX have been shown to bind the SH3 motif of the adaptor molecule Nck (Su, Y-C. et al, EMBO J. 16, 1279-1290 (1997)). From this evidence, human ZC1 may have the potential to bind to Nck or other SH3 or WW domain-containing proteins and participate in growth factor-induced signaling pathways.

The 256 amino acid spacer region of human ZC1 is most related to human STE20-subfamily kinases, ZC2 (SEQ ID NO:14) and ZC3 (SEQ ID NO:15), as well as to human PITSLRE (GB:U04824), sharing 59.9%, 33.1%, 29.6%, and 26.4% amino acid identity, respectively. It also shares 59.9% amino acid homology to murine NIK. The C. elegans homologue ZC504.4 has only limited sequence similarity in this spacer region.

The 343 amino acid C-terminal of human ZC1 is most related to human STE20-subfamily kinases, ZC3 (SEQ ID NO:15), ZC2 (SEQ ID NO:14), and ZC4 (SEQ ID NO:16), sharing 89.2%, 88.9%, and 42.3%, amino acid identity, respectively. The ZC1 C-terminal domain also shares 98.8% amino acid identity to murine NIK. The C. elegans homologue ZC504.4 also shares 68.7% amino acid identity with the C-tail of human ZC1. A lower, yet significant, homology is also evident to human KHS2 (SEQ ID NO:18), GCK (GB:U07349), and murine citron (GB:U07349) with 26.6%, 23.1% and 36.2% amino acid identity, respectively. GCK is a STE20-family kinase whose C-terminal domain has been shown to bind the small G-protein Rab8 (Ren, M. et al., Proc. Natl. Acad. Sci. 93, 5151-5155 (1996)). Citron is a non-kinase Rho-binding protein (Madaule, P. et al., FEBS Lett. 377, 243-238 (1995)).

The sequence similarity of the C-terminal region of ZC1 to proteins that have potential Rab- or Rho-binding domains suggests that ZC1 may signal through a small G-protein-dependant pathway.

Mammalian ZC2

The 4055 bp human ZC2 nucleotide sequence of the partial cDNA encodes a polypeptide of 1297 amino acids (SEQ ID NO:14) with a predicted molecular mass of 147,785 daltons. Analysis of the deduced amino acid sequence predicts ZC2 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain, however the cDNA clone lacks an initiating ATG, so the full extent of it N-terminus is not known. The N-terminally truncated ZC2 protein contains a 255 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 187 amino acid region predicted to form a coiled-coil structure, a 184 amino acid proline-rich region, a 328 amino acid spacer region, followed by a 343 amino acid C-terminal domain containing a potential Rab/Rho-binding region.

ZC2 is most closely related to the human STE20-subfamily kinases ZC3 (SEQ ID NO:15), ZC1 (SEQ ID NO:13), and ZC4 (SEQ ID NO:16), sharing 88.3%, 61.7%, and 41.9% amino acid identity, respectively, and shares 41.7% amino acid identity to a C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029).

The 255 amino acid catalytic domain of human ZC2 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15), SOK-1 (GB:X99325), KHS2 (SEQ ID NO:18), MST1 (GB:U18297), and GCK (GB:U07349), and to the C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029) sharing 90.2%, 89.8%, 49.0%, 48.6%, 47.9%, 45.0 and 76.7% amino acid identity, respectively. ZC2 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3, PAK4 and PAK5.

Immediately C-terminal to the kinase domain of human ZC2 is a 187 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm (supra). This region of ZC2 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15), and GEK2 (SEQ ID NO:107), as well as to human PITSLRE (GB:U04824), sharing 65.8%, 61.5%, 29.7% and 29.6% amino acid identity, respectively. The C. elegans homologue ZC504.4 shares 30.8% sequence identity over this region. Human ZC2 lacks the potential leucine zipper found in ZC1 as a consequence of a 29 amino acid deletion relative to ZC1 and ZC3.

The 184 amino acid proline-rich region of human ZC2 is most related to human STE20-subfamily kinases, ZC3 (SEQ ID NO:15) and ZC1 (SEQ ID NO:13), sharing 35.9% and 28.6%,amino acid identity, respectively. Significant homology is also evident to the murine WW domain-binding protein WBP7 (GB:U92455), and to the human SH3 domain-binding protein 3BP-1 (GB:X87671), with 27.7% and 25.3% amino acid identity, respectively.

ZC2 contains two of the potential “PxxP” (SEQ ID NO: 148) SH3 domain-binding motifs (II and III) found within the proline-rich region of human ZC 1. Motif II is conserved in ZC1, ZC3, ZC4 and C. elegans ZC504.4, and Motif III is conserved in ZC1, ZC3 and ZC4. Motifs II and III of murine NIX have been shown to bind the SH3 motif of the adaptor molecule Nck. From this evidence, human ZC1 may have the potential to bind to Nck or other SH3 or WW domain-containing proteins, and to participate in growth factor-induced signaling pathways.

The 328 amino acid spacer region of human ZC2 is most related to human STE20-subfamily kinases ZC1 (SEQ ID NO:13) and ZC3 (SEQ ID NO:15), and to murine NIK (GB:U88984), sharing 31.6%, 26.9% and 25.9% amino acid identity, respectively. The C. elegans homologue ZC504.4 has only limited sequence similarity in this spacer region.

The 343 amino acid C-terminal of human ZC2 is most related to human STE20-subfamily kinases ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15) and ZC4 (SEQ ID NO:16), and to murine NIK (GB:U88984), sharing 88.9%, 88.3%, 41.9%, and 88.0%, amino acid identity, respectively. The C. elegans homologue, ZC504.4, also shares 67.2% amino acid identity with the C-tail of human ZC2. A lower, yet significant, homology is also evident to human GCK (GB:U07349), murine citron (GB:U07349), and the S. cerevisiae ROM2 protein (GB:U19103), a Rhol GDP/GTP exchange factor, with 22.3%, 22.2% and 21.9% amino acid identity, respectively.

The sequence similarity of the C-terminal region of ZC2 to proteins that have potential Rab- or Rho-binding domains suggests that ZC2, like ZC1, may also signal through a small G-protein-dependant pathway.

Mammalian ZC3

The 4133 bp human ZC3 nucleotide sequence of the partial cDNA encodes a polypeptide of 1326 amino acids (SEQ ID NO:15) with a predicted molecular mass of 149,906 daltons. Analysis of the deduced amino acid sequence predicts ZC3 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain, however the cDNA clone lacks an initiating ATG, so the full extent of it N-termius is not known. The N-terminally truncated ZC3 protein contains a 255 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase: a 221 amino acid region predicted to form a coiled-coil structure, a 204 amino acid proline-rich region, and a 303 amino acid spacer region followed by a 343 amino acid C-terminal domain containing a potential Rab/Rho-binding region.

ZC3 is most closely related to the human STE20-subfamily kinases ZC1 (SEQ ID NO:13), ZC2 (SEQ ID NO:14), and ZC4 (SEQ ID NO:16), sharing 62.0%, 61.0%, and 42.5% amino acid identity, respectively and shares 46.7% amino acid identity to a C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029).

The 255 amino acid catalytic domain of human ZC3 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13), ZC2 (SEQ ID NO:14), SOK-1 (GB:X99325), KHS2 (SEQ ID NO:18), GCK (GB:U07349), SULU1 (SEQ ID NO:22), and GEK2 (SEQ ID NO:107), and to the C. elegans kinase encoded by the cosmid ZC504.4 (GB:Z50029) sharing 90.6%, 89.3%, 49.0%, 48.3%, 45.0%, 43.1%, 42.3% and 76.7% amino acid identity, respectively. ZC1 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC2, GEK2, KHS2, SULU1, SULU3, PAK4 and PAK5.

Immediately C-terminal to the kinase domain of human ZC3 is a 221 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm (supra). This region of ZC3 is most homologous to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13), ZC2 (SEQ ID NO:14), and GEK2 (SEQ ID NO:107), sharing 66.9%, 61.5%, and 27.5% identity, as well as to rat PLC-beta (GB:A45493) and human PITSLRE (GB:H54024) sharing 29.6% and 25.9% amino acid identity, respectively. The C. elegans homologue ZC504.4 shares 26.8% sequence identity over this region.

Within the predicted coiled-coil domain of human ZC3, and the related ZC1, is a region predicted to form a leucine zipper (Leu-X6-Leu-X6-Leu-X6-Leu-X20-Leu-X6-Leu) (SEQ ID NO: 149). The fact that this leucine repeat exists within a predicted coiled-coil structure suggests that the leucine zipper may have a high probability of serving as a dimerization interface (Hirst, J. D. et al Protein Engineering 9 657-662 (1996)) mediating a potential inter- or intra-molecular dimerization of human ZC3.

The 204 amino acid proline-rich region of human ZC3 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13) and ZC2 (SEQ ID NO:14), sharing 66.9% and 61.5% amino acid identity, respectively.

ZC3 contains two of the potential “PxxP” (SEQ ID NO: 148) SH3 domain-binding motifs (II and III) found within the proline-rich region of human ZC 1. Motif II is conserved in ZC1, ZC2, ZC4 and C. elegans ZC504.4; Motif III is conserved in ZC1, ZC2 and ZC4. Motifs II and III of murine NIK have been shown to bind the SH3 motif of the adaptor molecule Nck. From this evidence, human ZC3 may have the potential to bind to Nck or other SH3 or WW domain-containing proteins and participate in growth factor-induced signaling pathways.

The 303 amino acid acid spacer region of human ZC3 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13) and ZC2 (SEQ ID NO:14) sharing 30.1%, and 27.1% amino acid identity, respectively. The C. elegans homologue ZC504.4 lacks nearly the entire spacer region of ZC3.

The 343 amino acid C-terminal of human ZC3 is most related to human STE20-subfamily kinases, ZC1 (SEQ ID NO:13), ZC2 (SEQ ID NO:14) and ZC4 (SEQ ID NO:16), sharing 89.2%, 88.9%, and 42.5%, amino acid identity, respectively. The C. elegans homologue ZC504.4 also shares 67.2% amino acid identity with the C-tail of human ZC3. A lower, yet significant, homology is also evident to human GCK (GB:U07349), as well as to the non-kinases murine citron (GB:U07349) and the S. cerevisiae ROM2 protein (GB:U19103), a Rhol GDP/GTP exchange factor, with 21.6%, 32.4% and 22.9% amino acid identity, respectively.

The sequence similarity of the C-terminal region of ZC3 to proteins that have potential Rab- or Rho-binding domains suggests that ZC3, like ZC1 and ZC2, may signal through a small G-protein-dependant pathway.

Mammalian ZC4

The 3,684 bp human ZC4 nucleotide sequence of the complete cDNA encodes a polypeptide of 1,227 amino acids (SEQ ID NO:105) with a predicted molecular mass of 138,205 Daltons. Analysis of the deduced amino acid sequence predicts ZC4 to be an intracellular STE20-subfamily kinase, lacking both a signal sequence and a transmembrane domain. The full-length ZC4 protein contains a 25 amino acid N-terminus, a 265 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 108 amino acid region predicted to form a coiled-coil structure, a 231 amino acid proline-rich region, a 40 amino acid region predicted to form a coiled-coil structure spacer region, a 204 amino acid spacer region (domain B), followed by a 355 amino acid C-terminal domain containing a potential Rab/Rho-binding region (domain C).

ZC4 is most closely related to human ZC1 (SEQ ID NO:13, also known as human HGK, human KIAA0687, murine NIK, human AC005035, human NIK, and C. elegans MIG-15), ZC2 (SEQ ID NO:14, similar to partial sequence human KIAA0551), and ZC3 (SEQ ID NO:15). An assembled genomic fragment in the database (Z83850) is identical to ZC4, except for inappropriate identification of the exon boundaries. (Abo et al. (1998) EMBO J. 17: 6527-6540.)

The 25 amino acid N-terminal domain of human ZC4 shares weak homology to human ZC1 in its C-terminal extent, but otherwise does not reveal any significant homologies to the protein database.

The 265 amino acid catalytic domain of human ZC4 is most related to human ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15), and ZC2 (SEQ ID NO:14), sharing 63%, 64% and 62% amino acid identity, respectively.

Immediately C-terminal to the kinase domain of human ZC4 is a 108 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm (supra). This region is most related to human ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15), and ZC2 (SEQ ID NO:14), sharing 29%, 25% and 20% amino acid identity, respectively.

The 231 amino acid proline-rich region of human ZC4 does not reveal any significant homologies to the protein database. This region of ZC4 contains two “PxxP” (SEQ ID NO: 148) motifs that could potentially bind to proteins containing SH3 or WW domains and allow ZC4 to participate in growth factor activated signaling pathways. In addition, within the pro-rich domain of human ZC4 is a region predicted to form a leucine zipper (Leu-X6-Leu-X6-Leu-X6-Leu-X20-Leu-X6-Leu) (SEQ ID NO: 149) which may serve as a dimerization interface. The ZC STE2O subfamily kinases (ZC 1, ZC2 and ZC3) have similarly located “PxxP” (SEQ ID NO: 148) motifs and potential Leu zippers.

Immediately C-terminal to the proline-rich region of human ZC4 is a 40 amino acid region also predicted to form a coiled-coil structure based on the Lupas algorithm. This region of human ZC4 does not reveal any significant homologies to the protein database.

The 204 amino acid acidic- and serine-rich domain “B” of ZC4 does not reveal any significant homologies to the protein database.

The 355 amino acid C-terminal of human ZC4 is most related to human ZC1 (SEQ ID NO:13), ZC3 (SEQ ID NO:15), and ZC2 (SEQ ID NO:14), sharing 43%, 42% and 42% amino acid identity, respectively.

The sequence similarity of the C-terminal region of ZC4 to proteins that have potential Rab- or Rho-binding domains suggests that ZC4, like other ZC-subfamily STE20 kinases, may signal through a small G-protein-dependant pathway.

Mammalian KHS2

The 4023 bp human KHS2 nucleotide sequence encodes a polypeptide of 894 amino acids (SEQ ID NO:18) with a predicted molecular mass of 101,327 daltons. Analysis of the deduced amino acid sequence predicts KHS2 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The full-length KHS2 protein contains a 13 amino acid N-terminus, a 260 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 73 amino acid spacer region, a 188 proline-rich region, followed by a 360 amino acid C-terminal domain containing a potential Rab/Rho-binding site.

KHS2 is most closely related to the human STE20-subfamily kinases KHS1 (GB:U177129), GCK (GB:U07349), and HPKl (GB:U07349), sharing 65.5%, 51.9%, and 44.9% amino acid identity, respectively and shares 38.5% amino acid identity to a C. elegans STK (GB:U55363).

The 13 amino acid N-terminal domain of human KHS2 does not reveal any significant homologies that might suggest a potential function for this domain when examined by a Smith-Waterman alignment to the nonredundant protein database. Human KHS2 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristylation.

The 260 amino acid catalytic domain of human KHS2 is most related to human STE20-subfamily kinases KHS1 (GB:U177129), GCK (GB:U07349), HPK1 (GB:U66464), SOK-1 (GB:X99325), MST1 (GB:U18297), ZC1 (SEQ ID NO:13), and to the C. elegans kinase (GB:U55363), sharing 85.4%, 75.1%, 67.7%, 51.4%, 48.1%, 49.8% and 72.0% amino acid identity, respectively. KHS2 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC2, ZC3, ZC4, GEK2, SULU1, SULU3, PAK4 and PAK5.

The 73 amino acid acid spacer region of human KHS2 is most related to human STE20-subfamily kinases, KHS1 (GB:U177129), HPKl (GB:U66464) and GCK (GB:U07349), sharing 60.3%, 43.5% and 44.0%, amino acid identity, respectively.

The 188 amino acid proline-rich region of human KHS2 is most related to human STE20-subfamily kinases, HPK1 (GB:U66464), GCK (GB:U07349) and KHS1 (GB:U177129), sharing 33.3%, 31.9% and 31.4%, amino acid identity, respectively.

Two potential “PxxP” (SEQ ID NO: 148) SH3 domain-binding motifs (I and II) are found within the proline-rich region of human KHS2. Motif I is conserved with human KLHSl and HPK1; motif II is conserved with GCK and KHS2. A 192 amino acid region of human HPK1 containing motif II has been shown to bind to the C-terminal SH3 motif of the adaptor molecule Grb2 (Anafi, M et al, J. Biol. Chem. J. 272, 27804-27811 (1997)). Human KHS2 may bind SH3 or WW domain-containing proteins through this proline-rich region.

The 360 amino acid C-terminal of human KHS2 is most related to KHS1 (GB:U177129), GCK (GB:U07349) and HPK1 (GB:U66464), and to the C. elegans kinase (GB:U55363), sharing 74.9%, 54.8%, 42.9%, and 31.0%, amino acid identity, respectively. GCK is a STE20-family kinase whose C-terminal domain has been shown to bind the small G-protein Rab8 (Ren, M. et al., Proc. Natl. Acad. Sci. 93, 5151-5155 (1996)).

Mammalian SULU1

The 4196 bp human SULU1 nucleotide sequence encodes a polypeptide of 898 amino acids (SEQ ID NO:22) with a predicted molecular mass of 105,402 daltons. Analysis of the deduced amino acid sequence predicts SULU1 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The full-length SULU1 protein contains a 21 amino acid N-terminus, a 256 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 150 amino acid spacer region, a 210 amino acid region predicted to form a coiled-coil structure, a 114 amino acid spacer region and a 147 amino acid C-terminal domain predicted to form a coiled-coil structure.

SULU1 is most closely related to the STE20-subfamily kinases murine SULU3 (SEQ ID NO:24), human SULU3 (SEQ ID NO:23), and to the C. elegans kinase SULU (GB:U11280), sharing 68.9%, 72.2% and 38.2% amino acid identity, respectively.

The 21 amino acid N-terminal domain of human SULU1 is most related to murine SULU3 (SEQ ID NO:24) and to the C. elegans kinase SULU (GB:U11280), sharing 86.3% and 62.3% amino acid identity. Human SULU1 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristoylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 256 amino acid catalytic domain of human SULU1 is most related to murine SULU3 (SEQ ID NO:24), and to human SOK-1 (GB:X99325), STLK2 (SEQ ID NO:5), MST1 (GB:U18297), PAK1 (GB:U24152), ZC2 (SEQ ID NO:14), and KHS2 (SEQ ID NO:18) sharing 86.3%, 48.1%, 46.9%, 45.2%, 43.3%, 43.1% and 42.0% amino acid identity, respectively. The C. elegans SULU STK (GB:U11280) shares 62.3% sequence identity over this region. SULU1 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC2, ZC3, ZC4, GEK2, KHS2, SULU3, PAK4 and PAK5.

The 150 amino acid spacer region of human SULU1 is most related to human SULU3 (SEQ ID NO:23) and to the C. elegans kinase (GB:U11280), sharing 53.5% and 10.4% amino acid identity, respectively.

Immediately C-terminal to the spacer region of human SULU1 is a 210 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of SULU1 is most related to SULU3 (SEQ ID NO:23), the C. elegans SULU kinase (GB:U11280), GEK 2 (SEQ ID NO:107) and ZC1 (SEQ ID NO:13), sharing 68.6%,26.8%,23.2%, and 22.8% amino acid identity, respectively.

The 114 amino acid spacer region human SULU1 is most related to human SULU3 (SEQ ID NO:24) with 73.7% amino acid sequence identity. A lower, yet significant, homology is also evident to murine PITSLRE (GB:U04824) and DLK (GB:A55318), human ZC1 (SEQ ID NO:13) and GEK 2 (SEQ ID NO:107), as well as to the C. elegans SULU STK (GB:U11280), sharing 39.7%, 35.4%, 29.5%, 23.6% and 37.6% amino acid identity, respectively.

Immediately C-terminal to the second spacer region of human SULU1 is a 147 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of SULU1 is most related to human SULU3 (SEQ ID NO:24), ZC1 (SEQ ID NO:13) and GEK 2 (SEQ ID NO:107), as well as to the C. elegans SULU STK (GB:U11280), sharing 73.3%, 28.4%, 26.1% and 39.5%, amino acid identity, respectively.

Mammalian (Human) SULU3

The 3824 bp partial cDNA human SULU3 nucleotide sequence encodes a polypeptide of 786 amino acids (SEQ ID NO:23) with a predicted molecular mass of 92,037 daltons. Analysis of the deduced amino acid sequence predicts SULU3 to be an intracellular serine/threonine kinase lacking a transmembrane domain. The N-terminally truncated human SULU3 protein contains a 66 amino acid partial catalytic domain followed by a 149 amino acid spacer region, a 210 amino acid region predicted to form a coiled-coil structure, a second spacer region of 114 amino acids, a 247 amino acid C-terminal region predicted to form a second coiled-coil structure and a 100 amino acid C-terminal tail.

Human SULU3 is most closely related murine SULU3 (SEQ ID NO:24), human SULU1 (SEQ ID NO:22), and to the C. elegans SULU kinase (GB:U11280), sharing 66.3%, 68.9% and 32.9% amino acid identity, respectively. The high sequence homology between murine and human SULU3 suggests that these two proteins are orthologs of each other.

The 66 amino acid partial catalytic domain of human SULU3 is most related to murine SULU3 (SEQ ID NO:24), and to the human STE20 subfamily kinases ZC1 (SEQ ID NO:13), STE20 (GB:X99325), KHS1(GB:U177129) and GEK 2 (SEQ ID NO:107), as well as to the C. elegans SULU kinase (GB:U11280), sharing 83.3%, 47.0%, 45.5%, 43.5%,41.8% and 55.6% amino acid identity, respectively.

The 149 amino acid spacer region of human SULU3 is most related to murine SULU3 (SEQ ID NO:24), human STE20 (GB:X99325), MST1 (GB:U18297), and to the C.elegans SULU kinase (GB:U11280) sharing 98.7%, 21.9% and 21.8% amino acid identity, respectively.

Immediately C-terminal to the first spacer region of human SULU3 is a 210 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of SULU3 is most related to murine SULU3 (SEQ ID NO:24), and to human SULU1 (SEQ ID NO:22), ZC1 (SEQ ID NO:13) and GEK 2 (SEQ ID NO:107), as well as to the C. elegans SULU kinase (GB:U11280), sharing 99.5%, 68.6%, 27.4% and 22.5% amino acid identity, respectively.

The 114 amino acid second spacer region of human SULU3 is most related to murine SULU3 (SEQ ID NO:24), and to human SULU1 (SEQ ID NO:22) GEK 2 (SEQ ID NO:107), and ZC1 (SEQ ID NO:13), as well as to the C. elegans SULU kinase (GB:U11280), sharing 99.1%, 73.7%, 24.6%,24.1% and 41.2% amino acid identity, respectively.

Immediately C-terminal to the second spacer region of human SULU3 is a 247 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm (supra). This region of SULU3 is most related to human SULU1 (SEQ ID NO:22) and ZC1 (SEQ ID NO:13) as well as to rat PKN-(GB:D26180) murine pl60 ROCK1 (GB:U58512), and the C. elegans SULU kinase (GB:U11280), sharing 73.7%, 26.7%, 24.0% and 21.0% amino acid identity, respectively.

The 100 amino acid C-tail of human SULU3 is most related to a human prion protein (GB:L38993), with 45.0% amino acid identity.

Mammalian (Murine) SULU3

The 2249 bp murine, partial cDNA SULU3 nucleotide sequence encodes a polypeptide of 748 amino acids (SEQ ID NO:24) with a predicted molecular mass of 87,520 daltons. Analysis of the deduced amino acid sequence predicts SULU3 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The partial murine SULU3 protein contains a 25 amino acid N-terminus, a 248 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 149 amino acid spacer region, a 210 amino acid region predicted to form a coiled-coil structure, and a 116 amino acid spacer region.

Murine SULU3 is most closely related to human SULU3 (SEQ ID NO:23) and SULU1 (SEQ ID NO:22), as well as to the C. elegans SULU kinase (GB:U112 80), sharing 97.0%, 72.3% and 38.4% amino acid identity, respectively. The high sequence homology between murine and human SULU3 suggests that these two proteins are orthologs.

The 25 amino acid N-terminal domain of murine SULU3 is most related to human SULU1 (SEQ ID NO:22) and to the C. elegans SULU kinase (GB:U11280), sharing 70.0% and 44.4% amino acid identity, respectively.

Murine SULU3 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristoylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 248 amino acid catalytic domain of murine SULU3 is most related to human SULU1 (SEQ ID NO:22), STE20 (GB:X99325), ZC1 (SEQ ID NO:13), and KHS1 (GB:U77129), as well as to the C. elegans SULU kinase (GB:U11280), sharing 86.7%, 46.6%, 43.3%, 59.4% amino acid identity, respectively. Murine SULU3 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3, PAK4 and PAK5.

The 149 amino acid spacer of murine SULU3 is most related to human SULU3 (SEQ ID NO:23), SULU1 (SEQ ID NO:22), and STE20 (GB:X99325), as well as to the C. elegans SULU (GB:U11280) and the S. cerevisiae STE20 (GB:L04655) kinases, sharing 98.7%, 53.4%, 21.9%, 59.4% and 21.9% amino acid identity, respectively.

Immediately C-terminal to the spacer region of murine SULU3 is a 210 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of murine SULU3 is most related to human SULU3 (SEQ ID NO:23), ZC1 (SEQ ID NO:13), and GEK 2 (SEQ ID NO:107), as well as to the C. elegans SULU kinase (GB:U11280), sharing 99.5%, 27.4%, 22.5% and 29.2% amino acid identity, respectively.

The 116 amino acid C-terminal spacer region of murine SULU3 is most related to human SULU3 (SEQ ID NO:23), GEK 2 (SEQ ID NO:107), and ZC1 (SEQ ID NO:13), well as to the C. elegans SULU kinase (GB:U11280), sharing 98.3%, 24.6%, 24.1% and 40.5% amino acid identity, respectively.

Mammalian (Murine/Human) SULU3

The 2249 bp murine SULU3 and the 3824 bp human SULU3 cDNAs contain a 1620 nucleotide overlap (541 amino acids) with 90% and 98% DNA and amino acid sequence identity, respectively. Owing to the high degree of sequence identity in this extended overlap, we propose that these are functional orthologues of a single gene. The combined murine/human 4492 bp SULU3 sequence encodes a polypeptide of 1001 amino acids (SEQ ID NO:31) with a predicted molecular mass of 116,069 daltons. Analysis of the deduced amino acid sequence predicts SULU3 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. SULU3 contains a 25 amino acid N-terminus, a 248 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 149 amino acid spacer region, a 210 amino acid region predicted to form a coiled-coil structure and a second spacer region of 114 amino acids, a 247 amino acid C-terminal region predicted to form a second coiled-coil structure and a 100 amino acid C-terminal tail. The murine SULU3 clone lacks the region from the second C-terminal coiled-coil to the C-terminus, whereas the human clone lacks the N-terminal domain, and all but 66 amino acids of the 248 amino acid kinase domain.

SULU3 is most closely related to SULU1 (SEQ ID NO:22) and the C. elegans SULU kinase (GB:U11280) sharing 72.3% and 38.4% amino acid identity, respectively.

The 25 amino acid N-terminal domain of SULU3 is most related to human SULU1 (SEQ ID NO:22) and to the C. elegans SULU kinase (GB:U11280), sharing 70.0% and 44.4% amino acid identity, respectively. SULU3 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 248 amino acid catalytic domain of SULU3 is most related to human SULU1 (SEQ ID NO:22), SOK-1 (GB:X99325), ZC1 (SEQ ID NO:13), KHS1 (GB:U77129) and the C. elegans SULU kinase (GB:U11280), sharing 86.7%, 46.6%, 43.3%, 42.0% and 59.4% amino acid identity, respectively. SULU3 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, PAK4 and PAK5.

The 149 amino acid spacer of SULU3 is most related to SULU1 (SEQ ID NO:22) and SOK-1 (GB:X99325), and to the C. elegans SULU (GB:U11280), and S. cerevisiae STE20 (GB:L04655) kinases, sharing 53.4%, 21.9%, 59.4% and 21.9% amino acid identity, respectively.

Immediately C-terminal to the spacer region of SULU3 is a 210 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region is most related to ZC1 (SEQ ID NO:13), GEK 2 (SEQ ID NO:107), and the C. elegans SULU kinase (GB:U11280), sharing 27.4% 22.5% and 29.2% amino acid identity, respectively.

The 114 amino acid spacer region of SULU3 is most related to human SULU1 (SEQ ID NO:22), GEK 2 (SEQ ID NO:107), ZC1 (SEQ ID NO:13), and to the C. elegans SULU kinase (GB:U11280), sharing 73.7%, 24.6%, 24.1% and 41.2% amino acid identity, respectively.

Immediately C-terminal to the second spacer region of SULU3 is a 247 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of SULU3 is most related to human SULU1 (SEQ ID NO:22) and ZC1 (SEQ ID NO:13), as well as to rat PKN (GB:D26180), murine p160 ROCK1 (GB:U58512) and the C. elegans SULU kinase (GB:U11280), sharing 73.7%, 26.7%, 24.0%, 21.0% and 37.6% amino acid identity, respectively.

The 100 amino acid C-tail of SULU3 is most related to a human prion protein (GB:L38993) with 45.0% amino acid identity.

Mammalian GEK2

The 2926 bp human GEK2 nucleotide sequence of the complete cDNA encodes a polypeptide of 968 amino acids (SEQ ID NO:107) with a predicted molecular mass of 112,120 daltons. Analysis of the deduced amino acid sequence predicts GEK2 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The complete GEK2 protein contains a 33 amino acid N-terminus, a 261 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase, a 43 amino acid spacer region, a 135 amino acid proline-rich region, a 252 amino acid region predicted to form a coiled-coil structure followed by a 244 amino acid region also predicted to form a coiled-coil structure.

GEK2 is most closely related to rat AT1-46 (GB:U33472) (a partial cDNA that extends from the middle of the first potential coiled-coil domain of GEK2 to the C-terminus), murine LOK (GB:D89728), Xenopus laevis polo-like kinase 1 (GB:AF100165), and human SLK (GB:AB002804), sharing 91.3%, 88.5%, 65.0%, and 44.7% amino acid identity, respectively. The high sequence homology between human GEK2, murine LOK and rat AT1-46 suggests that human GEK2 is a highly related protein to the rodent forms, or alternatively, its orthologue. Recently, a full-length version of GEK2 was reported (STK10 or human LOK AB015718). The 968 amino acid sequence is 99% identical to GEK2 (SEQ ID NO:107).

The 33 amino acid N-terminal domain of human GEK2 is most related to murine LOK (GB:D89728) and to human SLK (GB:AB002804), sharing 100% and 54.5% amino acid identity, respectively.

Human GEK2 lacks a glycine residue at position 2, and is therefore unlikely to undergo myristylation. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this domain.

The 261 amino acid catalytic domain of human GEK2 is most related to murine LOK (GB:D89728), rat AT1-46 (GB:D89728) and human SLK (GB:AB002804) as well as to a C. elegans kinase (GB:Z81460), sharing 97.7%, 90.8%, 54.5% and 55.9% amino acid identity, respectively. GEK2 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3, PAK4 and PAK5.

The 43 amino acid spacer region of human GEK2 is most related to murine LOK (GB:D89728) and to human SLK, sharing 83.7% and 77.6% amino acid identity, respectively.

The 135 amino acid proline-rich region of human GEK2 is most related to murine LOK (GB:D89728) with 66.2% amino acid identity, respectively. Within the proline-rich region of human GEK2 is a potential “PxxP” (SEQ ID NO: 148) SH3-binding domain conserved with murine LOK.

Immediately C-terminal to the proline-rich region of human GEK2 is a 252 amino acid region predicted to form a coiled-coil structure based on the Lupas algorithm. This region of human GEK2 is most related to rat AT1-46 (GB:D89728), murine LOK (GB:D89728) and human SLK (GB:AB002804), and ZC2 (SEQ ID NO:14), sharing 90.8%, 86.9%, 42.2%, and 29.7% amino acid identity, respectively.

Immediately C-terminal to the predicted coiled-coil structure of human GEK2 is a second potential coiled-coil structure of 244 amino acids predicted based on the Lupas algorithm. This region of human GEK2 is most related to rat AT1-46 (GB:D89728) and murine LOK (GB:D89728) as well as to human SLK (GB:AB002804) and ZC1 (SEQ ID NO:13), sharing 91.8%, 92.6%, 70.4% and 26.7% amino acid identity, respectively. The C. elegans kinase (GB:Z81460) shares 31.5% amino acid sequence identity over this region.

Mammalian PAK4

The 3604 bp human PAK4 nucleotide sequence encodes a polypeptide of 681 amino acids (SEQ ID NO:29) with a predicted molecular mass of 74,875 daltons. Analysis of the deduced amino acid sequence predicts PAK4 to be an intracellular serine/threonine kinase, lacking both a signal sequence and transmembrane domain. The full-length PAK4 protein contains a 51 amino acid N-terminus predicted to contain a rac-binding motif, a 173 amino acid insert relative to the known mammalian PAK proteins, a 169 amino acid spacer region, a 265 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase and a 23 amino acid C-terminal tail.

PAK4 is most closely related to human PAK5 (SEQ ID NO:30), PAK1 (GB: U24152), and PAK65 (GB:U25975), as well as to a C. elegans kinase (GB: Z74029), sharing 76.8%, 49.5%, 49.8%, and 34.6% amino acid identity, respectively.

The 51 amino acid N-terminal domain of human PAK4 is most related to human PAK1 (GB:U24152), and PAK65 (GB:U25975), as well as to a C.elegans kinase (GB: Z74029), sharing 50.0%, 50.0% and 49.0% amino acid identity, respectively.

The 10 amino acid region at positions 13-23 of human PAK4 fits the consensus for a Cdc42/Rac-binding motif (SXPX4-6HXXH) (SEQ ID NO: 150) (Burbelo, P.D., Dreschel, D. and Hall, A. J. Bio. Chem. 270, 29071-29074 (1995)).

The 173 amino acid insert of human PAK4, relative to the known mammalian PAK proteins, is most related to a C. elegans kinase (GB: Z74029) with 39.0% amino acid identity. A Smith-Waterman search of the nonredundant protein database does not reveal any significant homologies that might suggest a potential function for this region.

The 169 amino acid spacer of human PAK4 does not reveal any significant homologies that might suggest a potential function for this region. The equivalent spacer region in PAK1 binds to the guanine nucleotide exchange factor PIX (Manser, E. et al (1998) Molecular Cell, 1, 183-192). Since PAK4 differs substantially from PAK1 over this region, the spacer domain of PAK4 may differ in its guanine nucleotide exchange factor binding specificity, relative to PAK1.

The 265 amino acid catalytic domain of human PAK4 is most related to human PAK5 (SEQ ID NO:30), PAK1 (GB:U24152), GCK (GB:U07349), SOK-1 (GB:X99325), and SLK (GB:AB002804), as well as to the C. elegans (GB: Z74029), and S. cerevisiae STE20-related kinases (GB:L04655), sharing 95.9%, 51.7%, 41.3%, 39.8%, 37.4%, 60.2% and 47.9% amino acid identity, respectively. PAK4 contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3 and PAK5.

The 23 amino acid C-tail of human PAK4 contains a sequence that is homologous to a G-protein beta subunit binding site (Leeuw, T. et al. Nature, 391, 191-195 (1998)). PAK4 has, therefore, the potential to be activated by both Cdc42- as well as G-protein-dependant pathways.

Mammalian PAK5

The 2,806 bp human PAK5 nucleotide sequence of the complete cDNA encodes a polypeptide of 591 amino acids (SEQ ID NO:103) with a predicted molecular mass of 64,071 Daltons. Analysis of the deduced amino acid sequence predicts PAK5 to be an intracellular STE20-subfamily kinase, lacking both a signal sequence and transmembrane domain. The full-length PAKS protein contains a 52 amino acid N-terminus predicted to contain a p21 (small G-protein) binding domain (PDB or CRIB), a 121 amino acid-insert relative to the known mammalian PAK proteins, a 134 amino spacer region, a 265 amino acid catalytic domain with all the motifs characteristic of a serine/threonine kinase and a 19 amino acid C-terminal tail.

PAK5 is most closely related to Human PAK4 (SEQ ID NO:29), Drosophila melanogaster PAK (also known as “mushroom bodies tiny”) (AJ011578), C45B11.1b from C. elegans (Z74029), and human PAK3 (Q13177) sharing 48% (327/674 aa), 50% (330/651 aa), 43% (234/435 aa excluding gap), and 47% (190/405 aa excluding gap) amino acid identity, respectively. Recently, a full length version of PAK5 was reported (PAK4 AF005046) whose 591 amino acid sequence is identical to PAK5 (SEQ ID NO:103). (Abo, et al. (1998) EMBO J. 17:6527-6540).

The 52 amino acid N-terminal domain of human PAK5 is most related to human PAK4 (SEQ ID NO:29), Drosophila melanogaster PAK (AJ011578), C45B11.1b from C. elegans (Z74029), and human PAK3 (Q13177), sharing 65%, 57%, 54%, and 53% amino acid identity, respectively.

The 11 amino acid region at positions 12-22 of human PAK5 (FIG.10A) fits the consensus for a small G-protein binding domain (PDB or CRIiB) (SXPX4-6HXXH) (SEQ ID NO: 150) (Burbelo, P. D., Dreschel, D. and Hall, A. J. Bio. Chem. 270, 29071-29074 (1995), hereby incorporated by reference herein in its entirety including any figures, tables, or drawings.).

The 121 amino acid insert of human PAK5 shares 43% amino acid identity with a similar domain from PAK4 (SEQ ID NO:29), but that is absent from other known PAKs.

The equivalent spacer region in PAK1 binds to the guanine nucleotide exchange factor PIX (Manser, E. et al (1998) Molecular Cell, 1, 183-192 hereby incorporated by reference herein in its entirety including any drawings, figures, or tables.). Since PAK5 differs substantially from PAK1 over this region, the spacer domain of PAK5 may differ in its guanine nucleotide exchange factor binding specificity, relative to PAK1.

The 134 amino acid collagen-like region of human PAK5 shares 34% amino acid identity to pro-α I type collagen from several species and is not present in other known PAKs.

The 265 amino acid catalytic domain of human PAK5 is most related to human PAK4 (SEQ ID NO:29), Drosophila melanogaster PAK (AJ011578), C45B11.1b from C. elegans (Z74029), and human PAK3 (Q13177), sharing 78%, 80%, 61%, and 55% amino acid identity, respectively. PAK5 also contains the potential “TPY” regulatory phosphorylation site in its activation loop. This “TPY” motif is conserved in other STE20-related kinases, including ZC1, ZC2, ZC3, ZC4, GEK2, KHS2, SULU1, SULU3 and PAK4.

The 19 amino acid C-tail shares 80% amino acid identity to a PAK-like homologue identified from genomic DNA (AL031652). Furthermore, this C-terminal region of human PAK5 contains a sequence that is homologous to a G-protein beta subunit binding site (Leeuw, T. et al. Nature, 391, 191-195 (1998) hereby incorporated by reference herein in its entirety including any figures, tables, or drawings). PAK5 has, therefore, the potential to be activated by both, Cdc42 as well as G-protein-dependant pathways.

V. Antibodies, Hybridomas, Methods of Use and Kits for Detection of STE20-Related Kinases

The present invention relates to an antibody having binding affinity to a kinase of the invention. The polypeptide may have the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, or SEQ ID NO:107, or a functional derivative thereof, or at least 9 contiguous amino acids thereof (preferably, at least 20, 30, 35, or 40 or more contiguous amino acids thereof).

The present invention also relates to an antibody having specific binding affinity to a kinase of the invention. Such an antibody may be isolated by comparing its binding affinity to a kinase of the invention with its binding affinity to other polypeptides. Those which bind selectively to a kinase of the invention would be chosen for use in methods requiring a distinction between a kinase of the invention and other polypeptides. Such methods could include, but should not be limited to, the analysis of altered kinase expression in tissue containing other polypeptides.

The STE20-Related kinases of the present invention can be used in a variety of procedures and methods, such as for the generation of antibodies, for use in identifying pharmaceutical compositions, and for studying DNA/protein interaction.

The kinases of the present invention can be used to produce antibodies or hybridomas. One skilled in the art will recognize that if an antibody is desired, such a peptide could be generated as described herein and used as an immunogen. The antibodies of the present invention include monoclonal and polyclonal antibodies, as well fragments of these antibodies, and humanized forms. Humanized forms of the antibodies of the present invention may be generated using one of the procedures known in the art such as chimerization or CDR grafting.

The present invention also relates to a hybridoma which produces the above-described monoclonal antibody, or binding fragment thereof. A hybridoma is an immortalized cell line which is capable of secreting a specific monoclonal antibody.

In general, techniques for preparing monoclonal antibodies and hybridomas are well known in the art (Campbell, “Monoclonal Antibody Technology: Laboratory Techniques in Biochemistry and Molecular Biology,” Elsevier Science Publishers, Amsterdam, The Netherlands, 1984; St. Groth et al., J. Immunol. Methods 35:1-21, 1980). Any animal (mouse, rabbit, and the like) which is known to produce antibodies can be immunized with the selected polypeptide. Methods for immunization are well known in the art. Such methods include subcutaneous or intraperitoneal injection of the polypeptide. One skilled in the art will recognize that the amount of polypeptide used for immunization will vary based on the animal which is immunized, the antigenicity of the polypeptide and the site of injection.

The polypeptide may be modified or administered in an adjuvant in order to increase the peptide antigenicity. Methods of increasing the antigenicity of a polypeptide are well known in the art. Such procedures include coupling the antigen with a heterologous protein (such as globulin or β-galactosidase) or through the inclusion of an adjuvant during immunization.

For monoclonal antibodies, spleen cells from the immunized animals are removed, fused with myeloma cells, such as SP2/0-Agl4 myeloma cells, and allowed to become monoclonal antibody producing hybridoma cells. Any one of a number of methods well known in the art can be used to identify the hybridoma cell which produces an antibody with the desired characteristics. These include screening the hybridomas with an ELISA assay, western blot analysis, or radioimmunoassay (Lutz et al., Exp. Cell Res. 175:109-124, 1988). Hybridomas secreting the desired antibodies are cloned and the class and subclass are determined using procedures known in the art (Campbell, “Monoclonal Antibody Technology: Laboratory Techniques in Biochemistry and Molecular Biology”, supra, 1984).

For polyclonal antibodies, antibody-containing antisera is isolated from the immunized animal and is screened for the presence of antibodies with the desired specificity using one of the above-described procedures. The above-described antibodies may be detectably labeled. Antibodies can be detectably labeled through the use of radioisotopes, affinity labels (such as biotin, avidin, and the like), enzymatic labels (such as horse radish peroxidase, alkaline phosphatase, and the like) fluorescent labels (such as FITC or rhodamine, and the like), paramagnetic atoms, and the like. Procedures for accomplishing such labeling are well-known in the art, for example, see Stemberger et al., J. Histochem. Cytochem. 18:315, 1970; Bayer et al., Meth. Enzym. 62:308-, 1979; Engval et al., Immunol. 109:129-, 1972; Goding, J. Immunol. Meth. 13:215-, 1976. The labeled antibodies of the present invention can be used for in vitro, in vivo, and in situ assays to identify cells or tissues which express a specific peptide.

The above-described antibodies may also be immobilized on a solid support. Examples of such solid supports include plastics such as polycarbonate, complex carbohydrates such as agarose and sepharose, acrylic resins and such as polyacrylamide and latex beads. Techniques for coupling antibodies to such solid supports are well known in the art (Weir et al., “Handbook of Experimental Immunology” 4th Ed., Blackwell Scientific Publications, Oxford, England, Chapter 10, 1986; Jacoby et al., Meth. Enzym. 34, Academic Press, N.Y., 1974). The immobilized antibodies of the present invention can be used for in vitro, in vivo, and in situ assays as well as in immunochromotography.

Furthermore, one skilled in the art can readily adapt currently available procedures, as well as the techniques, methods and kits disclosed herein with regard to antibodies, to generate peptides capable of binding to a specific peptide sequence in order to generate rationally designed antipeptide peptides (Hurby et al., “Application of Synthetic Peptides: Antisense Peptides”, In Synthetic Peptides, A User's Guide, W.H. Freeman, NY, pp. 289-307, 1992; Kaspczak et al., Biochemistry 28:9230-9238, 1989).

Anti-peptide peptides can be generated by replacing the basic amino acid residues found in the peptide sequences of the kinases of the invention with acidic residues, while maintaining hydrophobic and uncharged polar groups. For example, lysine, arginine, and/or histidine residues are replaced with aspartic acid or glutamic acid and glutamic acid residues are replaced by lysine, arginine or histidine.

The present invention also encompasses a method of detecting a STE20-related kinase polypeptide in a sample, comprising: (a) contacting the sample with an above-described antibody, under conditions such that immunocomplexes form, and (b) detecting the presence of said antibody bound to the polypeptide. In detail, the methods comprise incubating a test sample with one or more of the antibodies of the present invention and assaying whether the antibody binds to the test sample. Altered levels of a kinase of the invention in a sample as compared to normal levels may indicate disease.

Conditions for incubating an antibody with a test sample vary. Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the antibody used in the assay. One skilled in the art will recognize that any one of the commonly available immunological assay formats (such as radioimmunoassays, enzyme-linked immunosorbent assays, diffusion based Ouchterlony, or rocket immunofluorescent assays) can readily be adapted to employ the antibodies of the present invention. Examples of such assays can be found in Chard (“An Introduction to Radioimmunoassay and Related Techniques” Elsevier Science Publishers, Amsterdam, The Netherlands, 1986), Bullock et al. (“Techniques in Immunocytochemistry,” Academic Press, Orlando, Fla. Vol. 1, 1982; Vol. 2, 1983; Vol. 3, 1985), Tijssen (“Practice and Theory of Enzyme Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology,” Elsevier Science Publishers, Amsterdam, The Netherlands, 1985).

The immunological assay test samples of the present invention include cells, protein or membrane extracts of cells, or biological fluids such as blood, serum, plasma, or urine. The test samples used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed. Methods for preparing protein extracts or membrane extracts of cells are well known in the art and can be readily be adapted in order to obtain a sample which is testable with the system utilized.

A kit contains all the necessary reagents to carry out the previously described methods of detection. The kit may comprise: (i) a first container means containing an above-described antibody, and (ii) second container means containing a conjugate comprising a binding partner of the antibody and a label. In another preferred embodiment, the kit further comprises one or more other containers comprising one or more of the following: wash reagents and reagents capable of detecting the presence of bound antibodies.

Examples of detection reagents include, but are not limited to, labeled secondary antibodies, or in the alternative, if the primary antibody is labeled, the chromophoric, enzymatic, or antibody binding reagents which are capable of reacting with the labeled antibody. The compartmentalized kit may be as described above for nucleic acid probe kits. One skilled in the art will readily recognize that the antibodies described in the present invention can readily be incorporated into one of the established kit formats which are well known in the art.

VI. Isolation of Compounds Which Interact With STE20-Related Kinases

The present invention also relates to a method of detecting a compound capable of binding to a STE20-related kinase of the invention comprising incubating the compound with a kinase of the invention and detecting the presence of the compound bound to the kinase. The compound may be present within a complex mixture, for example, serum, body fluid, or cell extracts.

The present invention also relates to a method of detecting an agonist or antagonist of kinase activity or kinase binding partner activity comprising incubating cells that produce a kinase of the invention in the presence of a compound and detecting changes in the level of kinase activity or kinase binding partner activity. The compounds thus identified would produce a change in activity indicative of the presence of the compound. The compound may be present within a complex mixture, for example, serum, body fluid, or cell extracts. Once the compound is identified it can be isolated using techniques well known in the art.

The present invention also encompasses a method of agonizing (stimulating) or antagonizing kinase associated activity in a mammal comprising administering to said mammal an agonist or antagonist to a kinase of the invention in an amount sufficient to effect said agonism or antagonism. A method of treating diseases in a mammal with an agonist or antagonist of STE20-related kinase activity comprising administering the agonist or antagonist to a mammal in an amount sufficient to agonize or antagonize STE20-related kinase associated functions is also encompassed in the present application.

In an effort to discover novel treatments for diseases, biomedical researchers and chemists have designed, synthesized, and tested molecules that inhibit the function of protein kinases. Some small organic molecules form a class of compounds that modulate the function of protein kinases. Examples of molecules that have been reported to inhibit the function of protein kinases include, but are not limited to, bis monocyclic, bicyclic or heterocyclic aryl compounds (PCT WO 92/20642, published Nov. 26, 1992 by Maguire et al.), vinylene-azaindole derivatives (PCT WO 94/14808, published Jul. 7, 1994 by Ballinari et al.), 1-cyclopropyl-4-pyridyl-quinolones (U.S. Pat. No. 5,330,992), styryl compounds (U.S. Pat. No. 5,217,999), styryl-substituted pyridyl compounds (U.S. Pat. No. 5,302,606), certain quinazoline derivatives (EP Application No. 0 566 266 A1), seleoindoles and selenides (PCT WO 94/03427, published Feb. 17, 1994 by Denny et al.), tricyclic polyhydroxylic compounds (PCT WO 92/21660, published Dec. 10, 1992 by Dow), and benzylphosphonic acid compounds (PCT WO 91/15495, published Oct. 17, 1991 by Dow et al).

Compounds that can traverse cell membranes and are resistant to acid hydrolysis are potentially advantageous as therapeutics as they can become highly bioavailable after being administered orally to patients. However, many of these protein kinase inhibitors only weakly inhibit the function of protein kinases. In addition, many inhibit a variety of protein kinases and will cause multiple side-effects as therapeutics for diseases.

Some indolinone compounds, however, form classes of acid resistant and membrane permeable organic molecules. WO 96/22976 (published Aug. 1, 1996 by Ballinari et al.) describes hydrosoluble indolinone compounds that harbor tetralin, naphthalene, quinoline, and indole substituents fused to the oxindole ring. These bicyclic substituents are in turn substituted with polar moieties including hydroxylated alkyl, phosphate, and ether moieties. U.S. patent application Ser. No. 08/702,232, filed Aug. 23, 1996, entitled “Indolinone Combinatorial Libraries and Related Products and Methods for the Treatment of Disease” by Tang et al. (Lyon & Lyon Docket No. 221/187) and Ser. No. 08/485,323, filed Jun. 7, 1995, entitled “Benzylidene-Z-Indoline Compounds for the Treatment of Disease” by Tang et al. (Lyon & Lyon Docket No. 223/298) and International Patent Publication WO 96/22976, published Aug. 1, 1996 by Ballinari et al., all of which are incorporated herein by reference in their entirety, including any drawings, describe indolinone chemical libraries of indolinone compounds harboring other bicyclic moieties as well as monocyclic moieties fused to the oxindole ring. application Ser. No. 08/702,232, filed Aug. 23, 1996, entitled “Indolinone Combinatorial Libraries and Related Products and Methods for the Treatment of Disease” by Tang et al. (Lyon & Lyon Docket No. 221/187), Ser. No. 08/485,323, filed Jun. 7, 1995, entitled “Benzylidene-Z-Indoline Compounds for the Treatment of Disease” by Tang et al. (Lyon & Lyon Docket No. 223/298), and WO 96/22976, published Aug. 1, 1996 by Ballinari et al. teach methods of indolinone synthesis, methods of testing the biological activity of indolinone compounds in cells, and inhibition patterns of indolinone derivatives.

Other examples of substances capable of modulating kinase activity include, but are not limited to, tyrphostins, quinazolines, quinoxolines, and quinolines. The quinazolines, tyrphostins, quinolines, and quinoxolines referred to above include well known compounds such as those described in the literature. For example, representative publications describing quinazolines include Barker et al., EPO Publication No. 0 520 722 A1; Jones et al., U.S. Pat. No. 4,447,608; Kabbe et al., U.S. Pat. No. 4,757,072; Kaul and Vougioukas, U.S. Pat. No. 5,316,553; Kreighbaum and Corner, U.S. Pat. No. 4,343,940; Pegg and Wardleworth, EPO Publication No. 0 562 734 A1; Barker et al., Proc. of Am. Assoc. for Cancer Research 32:327 (1991); Bertino, J. R., Cancer Research 3:293-304 (1979); Bertino, J. R., Cancer Research 9(2 part 1):293-304 (1979); Curtin et al., Br. J. Cancer 53:361-368 (1986); Fernandes et al., Cancer Research 43:1117-1123 (1983); Ferris et al. J. Org. Chem. 44(2):173-178; Fry et al., Science 265:1093-1095 (1994); Jackman et al., Cancer Research 51:5579-5586 (1981); Jones et al. J. Med. Chem. 29(6):1114-1118; Lee and Skibo, Biochemistry 26(23):7355-7362 (1987); Lemus et al., J. Org. Chem. 54:3511-3518 (1989); Ley and Seng, Synthesis 1975:415-522 (1975); Maxwell et al., Magnetic Resonance in Medicine 17:189-196 (1991); Mini et al., Cancer Research 45:325-330 (1985); Phillips and Castle, J. Heterocyclic Chem. 17(19):1489-1596 (1980); Reece et al., Cancer Research 47(11):2996-2999 (1977); Sculier et al., Cancer Immunol. and Immunother. 23:A65 (1986); Sikora et al., Cancer Letters 23:289-295 (1984); Sikora et al., Analytical Biochem. 172:344-355 (1988); all of which are incorporated herein by reference in their entirety, including any drawings.

Quinoxaline is described in Kaul and Vougioukas, U.S. Pat. No. 5,316,553, incorporated herein by reference in its entirety, including any drawings.

Quinolines are described in Dolle et al., J. Med. Chem. 37:2627-2629 (1994); MaGuire, J. Med. Chem. 37:2129-2131 (1994); Burke et al., J. Med. Chem. 36:425-432 (1993); and Burke et al. BioOrganic Med. Chem. Letters 2:1771-1774 (1992), all of which are incorporated by reference in their entirety, including any drawings.

Tyrphostins are described in Allen et al., Clin. Exp. Immunol. 91:141-156 (1993); Anafi et al., Blood 82:12:3524-3529 (1993); Baker et al., J. Cell Sci. 102:543-555 (1992); Bilder et al., Amer. Physiol. Soc. pp. 6363-6143:C721-C730 (1991); Brunton et al., Proceedings of Amer. Assoc. Cancer Rsch. 33:558 (1992); Bryckaert et al., Experimental Cell Research 199:255-261 (1992); Dong et al., J. Leukocyte Biology 53:53-60 (1993); Dong et al., J. Immunol. 151(5):2717-2724 (1993); Gazit et al., J. Med. Chem. 32:2344-2352 (1989); Gazit et al., “J. Med. Chem. 36:3556-3564 (1993); Kaur et al., Anti-Cancer Drugs 5:213-222 (1994); Kaur et al., King et al., Biochem. J. 275:413-418 (1991); Kuo et al., Cancer Letters 74:197-202 (1993); Levitzki, A., The FASEB J. 6:3275-3282 (1992); Lyall et al., J. Biol. Chem. 264:14503-14509 (1989); Peterson et al., The Prostate 22:335-345 (1993); Pillemer et al., Int. J. Cancer 50:80-85 (1992); Posner et al., Molecular Pharmacology 45:673-683 (1993); Rendu et al., Biol. Pharmacology 44(5):881-888 (1992); Sauro and Thomas, Life Sciences 53:371-376 (1993); Sauro and Thomas, J. Pharm. and Experimental Therapeutics 267(3):119-1125 (1993); Wolbring et al., J. Biol. Chem. 269(36):22470-22472 (1994); and Yoneda et al., Cancer Research 51:4430-4435 (1991); all of which are incorporated herein by reference in their entirety, including any drawings.

Other compounds that could be used as modulators include oxindolinones such as those described in U.S. patent application Ser. No. 08/702,232 filed Aug. 23, 1996, incorporated herein by reference in its entirety, including any drawings.

VII. Biological Significance, Applications and Clinical Relevance of Novel STE20-Related Kinases Human STLK2, STLK3, STLK4, STLK5, STLK6, and STLK7

STLK2, STLK4, STLK5, STLK6 and STLK7 belong to an expanding family of intracellular STKs that have varying degrees of sequence homology to SOK-1, a kinase implicated in oxidative stress agents (Pombo, CM et al, EMBO J. (17) 4537-4546, 1996). Our data shows that STLK2 is expressed highly in hematopoietic cells. Therefore, STLK2 may participate in the oxidative response pathway during inflammation. In addition, STLK2 could also be a possible component in the signaling pathways leading to T cell activation. High levels of STLK2 in several tumor cell lines could also imply that STLK2 might be involved in tumorigenesis.

STLK2 is most closely related to two human STE20-subfamily kinases: MST3 and SOK-1. MST3 is a 52,000 daltons cytoplasmic kinase that is ubiquitously expressed with its highest levels of expression found in heart, skeletal muscle and pancreas. The serine/threonine kinase activity of MST3 is activated by phosphorylation. Unlike SOK-1, MST3 prefers Mn++over Mg++and can use both GTP and ATP as phosphate donors. MST3 may undergo dimerization. No agonists have yet been identified that activate MST3. The downstream signaling mechanism of this kinase is unknown (Schinkmann, K and Blenis, J. (1997) J. Biol. Chem. 272, 28695-28703).

SOK-1 is a 50,000 daltons cytoplasmic kinase expressed predominantly in testis, large intestine, brain and stomach and to a lesser extent in heart and lung. SOK-1 is also expressed in the germinal center B-cell line (RAMOS) and in a mature B cell line (HS Sultan). The serine/threonine kinase activity of SOK-1 is activated by phosphorylation. The C-terminus of SOK-1 has been shown to be inhibitory to the catalytic activity of this kinase. The only agonists known to activate SOK-1 are oxidant agents, like H₂O₂ and menadione, a quinone that is a potent intracellular generator of reactive oxygen species (Pombo, C. M. et al. EMBO J. 15, 4537-4546). SOK-1 is also activated by chemical anoxia through the generation of reactive oxygen species and release of calcium into the cytoplasm from intracellular stores. SOK-1, therefore, may play an important role in ischemia, the cause of myocardial infarction, stroke and acute renal failure (Pombo, C. M. et al. J. Biol. Chem. 272, 29372-29379 (1997)). The activity of SOK-1 in the response to oxidant stress is inversely correlated with the activity of the stress-activated protein kinases (SAPKs) elevated SOK-1 activity correlates with absent SAPK activity and vice-versa. SOK-1 does not activate any of the four MAP kinase pathways, SAPKs, p38, ERK-1 or MEK-5/ERK-5 (Pombo, C. M. et al. EMBO J. 15, 4537-4546). The downstream signaling mechanism of this kinase remains unknown.

STLK2 is expressed in a wide variety of immune cell types and tissues including thymus, dendrocytes, mast cells, monocytes, B cells (primary, Jurkat, RPMI, SR), T cells (CD8/CD4+, TH1, TH2, CEM, MOLT4) and megakaryocytes (K562), whereas STLK3 is restricted to thymus and STLK4 is predominately expressed in thymus, T cells (CD4/CD8+, TH1, CEM) and B cells (Jurkat, RPMI). Consequently, these STKs might participate in the oxidative response pathway during inflammation, reperfusion injury (stroke, surgery, shock), TNFa-mediated signaling, insulin desensitization, atherogenesis, vascular injury, T or B cell costimulation, or alternatively, participate in other MAPK-related signal transduction processes.

STLK5 is more distantly related to this STE20-subfamily including SOK-1 and STLK2, STLK3 and STLK4. STLK5, may therefore mediate a signaling pathway that is distinct from the oxidative stress response pathway.

The high degree of sequence homology in the C-termini of SOK-1, STLK2, STLK3, STLK4, STLK5, and STLK6 raises the possibility that these novel STKs, like SOK-1, may be subject to autoinhibition through a conserved C-terminal motif.

Human ZC1, ZC2, ZC3 and ZC4

ZC1 is a good candidate for any disease in which tyrosine kinase, cytokine, or heterotrimeric G-protein coupled receptors have been implicated. The mouse homologue binds to NCK, and is recruited to activated PDGF (Su et al., EMBO 16: 1279-1290, 1997). The Drosophila homolog has been shown to bind to TRAF2, implicating it in TNF-α signaling (Liu et al., (1999) Curr. Biol. 9:101-104, 1999)). While ZC1 does not contain the exact NCK- and TRAF2-binding domains, it is likely to bind to related proteins.

Of the ZC subfamily of STE20-related protein kinases, ZC1 has very broad over-expression in many tumor types, suggesting that it may be involved in cellular growth, transformation, or tumor progression. A truncated form of ZC1 containing only the C-terminal putative MEKK1-binding domain was found to reduce the number of foci generated by H-Ras-V12 in Rat Intestinal Epithelial cells (RIE-1). These data indicate that ZC1 may play a role in the ability for these cells to overcome contact inhibition and anchorage-dependent growth.

The ZC1 homolog, Misshapen (msn) in Drosophila melanogaster was cloned as a result of complementing a mutation in a developmental pathway required for dorsal closure, a process involving changes in cell shape and position in the embryo (Treisman et al. Gene 186 119-125, 1997). A D. melanogaster homolog of the JNK1/JNK2 kinases from mammals was shown to function downstream of msn in the dorsal-closure signaling pathway (Su et al. Genes Dev. 12:2371-2380, 1998).

While ZC1 could be involved in multiple aspects of tumorigenesis, by analogy with Drosophila, the role of misshapen in dorsal closure suggests a critical role in the regulation of the cytoskeleton for the processes of cell attachment, cell movement and perhaps migration.

The association of the ZC1 family members msn and NIK with TRAF2 may indicate a role for this kinase in cell survival and/or in apoptosis. The ZC1 family contains a highly conserved domain that in the mouse homolog, NIK, has been shown to bind to MEKK1 (Mitogen-activated/Extracellular-regulated Kinase Kinase 1) (Su et al., (1997) EMBO 16(6): 1279-90). MEKK1 is involved in cell survival and/or apoptosis in several systems (Schlesinger et al., Front. Biosci.3:D1181-6, 1998). Depending on the context, MEKK1 appears to be upstream of either the ERK1/MAPK or the JNK/SAPK pathway [Schlesinger et al., (1998 Front. Biosci. 3:D1181-6). Three homologues of ZC1: murine NIK (NCK-interacting kinase)(Su et al. EMBO 16:1279-90, 1997), Drosophila msn (Liu et al. Curr. Biol. 9:101-104, 1999) and human HGK (HPK/GCK-like kinase)(Yao et al., J. Biol. Chem. 274:2118-25, 1999) have all been shown to activate the JNK pathway when over-expressed in 293T cells.

ZC1 shares a high degree of homology with these other family members in both the kinase domain and the “MEKK”-binding domains, yet it differs in the intervening region, which contains several putative binding domains for upstream signaling adapter molecules (e.g. NCK, TRAF2). Unlike the other family members, ZC1 does not appear to activate the JNK pathway in 293T cells as seen by its ability to induce expression of either a JUN or ATF2-driven luciferase gene. Upon co-transfection into these cells with HA-tagged JNK, modest activation of JNK was detected. ZC1 also modestly activated co-transfected ERKI. Both the ERK and the JNK activation were very slight compared with the positive controls in the assay (activated forms of MEK1 and MEKK1, respectively). In both cases, activation required the full-length kinase. While the kinase domain alone is up to 5× more active in autophosphorylation and in phosphorylation of MBP, it does not lead to activation of these potential downstream kinases. Based on the strong sequence homology of ZC1 with the other family members, it is very likely that ZC1 will be important for either JNK or ERK activation once the proper context is found.

ZC1 profoundly inhibits ERK1 kinase expression in co-transfection assays. This effect is dependent on ZC1 kinase activity, occurring with the wild-type and the kinase domain alone, but not with the kinase-dead mutant even though all three forms of ZC1 are expressed at similar levels. This may suggest a role for this kinase in transcriptional or post-transcriptional regulation.

ZC1 may be an important component in the signaling pathways mediated by the co-stimulatory receptor CD28 in T cells and/or by the pro-inflammatory cytokine TNFα, since co-transfection of the wild-type ZC1 activated the RE/AP-luciferase and NFκB-luciferase reporter genes. While our data showed that ZC1 strongly activates NFθB in T-cells, no activation of NFκB driven luciferase was detectable in NIH 3T3 cells. A recent paper (J. Biol. Chem. 274:2118-25; 1999.) has shown that a human ZC1 splicing isoform, HGK, is involved in the TNFα-signaling pathways.

Given the importance of T cell activation in autoimmunity and transplantation, as well as the key role that TNFα plays in inflammatory diseases, it is possible that ZC1 could be a therapeutic target for immunological diseases which include but are not limited to: rheumatoid arthritus, chronic inflammatory bowel diseases (ie Crohn's disease), chronic inflammatory pelvic disease, multiple sclerosis, asthma, osteoarthritis, psoriasis, atherosclerosis, rhinitis, and autoimmunity as well as organ transplantation and cardiovascular diseases.

ZC1 appears to be the human orthologue of murine NIK and possibly an orthologue of a C. elegans STE20-subfamily kinase encoded by the ZC504.4 cosmid.

Murine NIIK is a 140,000 daltons kinase that is most highly expressed in brain and heart. NIK interacts with the SH3 domains of the adaptor molecule Nck through its proline-rich regions found in the C-terminal extra-catalytic region. The specific regions that mediate this interaction are two PxxP (SEQ ID NO: 148) motifs that are nearly uniformly conserved between NIX, ZC 1,2,3 and the C. elegans STE2O ZC504.4 kinase. In addition, NIX binds MEKK1 through its 719 amino acid C-terminal (Su, Y-C. et al. (1997) EMBO J. 16, 1279-1290). MEKK1 is a membrane-associated kinase responsible for activating MKK4 (also known as SEKi), which in turn activates SAPK (Yan, M et al. (1994) Nature, 372, 798-800). NIX may function as a kinase that links growth factor activated pathways and the stress-response pathway mediated by SAPKs. According to this hypothesis, activation of growth factor receptors leads to receptor tyrosine phosphorylation, Nck binding to the phosphorylated tyrosines via its SH2 domain, NIX redistribution to a membrane compartment via binding to the SH3 domain of Nck, and juxtaposition to the membrane-associated MEKK1. The NIX-MEKIKi interaction would, in this fashion, turn on the SAPK pathway in response to growth factor stimulation (Su, Y-C. et al. (1997) EMBO J. 16, 1279-1290).

Given the high homology between ZC1, ZC2, ZC3, and ZC4 STKs and NIK, it is conceivable that these kinases may each function to connect growth factor- and stress-activated signaling pathways. The heterogeneity that the ZC kinases exhibit within their putative SH3-binding domain could provide signaling specificity in terms of the nature of the adaptor molecule that they bind. The high level of sequence conservation in the C-termini of the ZC1, ZC2 and ZC3 strongly suggests that these human kinases, like murine NIK, also may bind to MEKK1 and activate SAPKs. The ZC kinases also display strong homology at their C-termini to protein domains that bind small GTPase proteins such as Rab, Rho and Rac. For example, the C-termini of ZC1 is 36.2% identical to citron, a murine Rho-binding protein, and 23.1% identical to the rab-binding region of GC kinase. This suggests that, in addition to adaptor molecules, small GTPase proteins may also mediate membrane association and activation of the ZC kinases. The presence of a potential coiled-coil region located immediately C-terminal to the catalytic region strongly suggests that the ZC kinases may also be subject to regulation via homo or heterodimerization events.

The C. elegans STE20 ZC504.4 kinase is the product of the mig-15 gene. The product of this gene has been implicated in several developmental processes such as epidermal development, Q neuroblast migrations and muscle arm targeting in the developing worm (Zhu, X. and Hedgecock E. (1997) Worm Breeder's Gazette 14, 76). The high level of sequence conservation between the ZC kinases and the ZC504.4 C. elegans kinase will make C. elegans a valuable model organism to study, through epistatic analysis, the signaling properties of the human ZC kinases.

Human KHS2

KHS1 (kinase homologous to SPS1/STE20) is a 100,000 dalton cytoplasmic STK that is expressed ubiquitously. KHS1 has been implicated in the mechanism of SAPK activation in response to inflammatory cytokines such as TNFα as well as to ultraviolight light, which also uses the TNF signaling pathway. TNFα binding to its receptors (TNFR1 and TNFR2) results in the sequential association with the receptor C-tail of multiple signaling molecules including TNFR1-associated death domain protein (TRADD), Fas-associated death domain protein (FADD or MORT1), TNFR-associated factor 2 (TRAF2), and the STK RIP (receptor interacting protein). The TRADD-TRAF2 interaction is mediated by a conserved region present at the C-terminus of TRAF2, the TRAF domain. Activation of the NFθB and SAPK pathways is mediated by the ring finger motif present at the N-terminus of TRAF2 (Curr. Opinion in Cell. Biol. (1997) 9:247-251). KHS1 is activated by TNFα stimulation in a TRAF2-dependant manner and inhibition of KHS1 blocks TNFα-induced SAPK activation but not NFκB activation. The mechanism by which TRAF2 activates KHS1 is not known. Cotransfection of TRAF2- and KHS1-expressing constructs in 293T cells failed to reveal a direct association between these two molecules. KHS1 activates the SAPK pathway by a direct association with the constitutively active kinase MEKK1. MEKK1 subsequently activates SEK1, which in turn activates SAPK. Neither the MAPK nor the p38 kinase pathways are activated by KHS1 (Shi, C-S and Kehrl. J. H. (1997) J. Biol. Chem. 272, 32102-32107). In addition to its catalytic domain, downstream signaling of KHS1 requires its conserved C-terminus (Diener, K. et al (1997) Proc. Natl. Acad. Sci. 94, 9687-9692).

GCK (germinal center kinase) is a constitutively active 97,000 dalton STK that is broadly expressed. GCK may participate in B-cell differentiation since its expression is localized to the germinal center within lymphoid follicles. GCK activates the SAPK pathway in response to TNFα via activation of SEK1. The upstream activators of GCK in response to cytokines as well as the immediate downstream target of this kinase are unknown. The C-terminus of GCK is sufficient to activate SEK1 (Pombo, C. M. et al (1995) Nature, 377, 750-754).

The murine orthologue of GCK, rab8ip (rab8-interacting protein), is a 97,000 dalton protein that fractionates with both the soluble cytoplasmic fraction as well as with a salt-sensitive fraction associated with the basolateral membrane of the trans-Golgi region in polarized MDCK epithelial cells. The C-terminus of rab8ip binds to rab8, a small GTP-binding protein required for vesicular transport from the Golgi apparatus (Ren, M. et al. (1996) Proc. Natl. Acad. Sci. 93, 5151-5155). In addition to inducing the transcriptional activation of cytokines like IL2 via SAPK, GCK may also promote the rab-dependent release of secretory proteins in response to TNFα (Buccione, R. et al (1995) Mol. Bio. Cell 6, 291).

HPK1 (hematopoietic protein kinase) is a constitutively active 90,000 dalton STK restricted to hematopoietic cells. HPKl activates the SAPK pathway by directly binding to and activating MEKK1 (Hu, M. et al (1996) Genes and Dev. 10:2251-2264) as well as the ubiquitously expressed mixed-lineage kinase MLK-3 (Kiefer, F. et al (1996) EMBO J. 15:7013-7025). This function of HPK1 requires, in contrast to GCK, both its kinase domain as well as its C-terminus. The upstream activators of HPK1 remain unknown. HPK1 also plays a key role as a mediator of transforming growth factor-β (TGFβ) signaling. HPK1 activates the TGFb-activated kinase (TAK), which in turn stimulates the SAPK pathway by phosphorylating SEK1 (Wang W. et al (1997) J. Biol. Chem. 272:22771-22775).

KHS2 is expressed in thymus, dendrocytes and monocytes. KHS2 could have a complementary function to that of KHS1 as a mediator of SAPK activation in the cellular response to inflammatory cytokines. KHS2 could have the potential to interact directly with TRAF2 since a STK with the predicted molecular weight of KHS2 (approximately 101,000 daltons) is found in the TNFR-TRAF2 complex upon TNFα stimulation (VanArsdale, T. and Ware, C. F. (1994) J. Immunol. 153, 3043-3050). The presence of a putative binding domain for Rab or a Rab-like molecule at the C-terminus of KHS2 indicates that KHS2, in addition to having a potential role in the TRAF2-dependant TNFα cytokine response, could also mediate signaling events that utilize small GTPase proteins. Alternatively, the binding of a small GTPase protein to the C-terminus of KHS2 may be required for its potential TRAF2-dependant signaling to a downstream kinase such as MEKK1.

Human GEK2, SULU1 and SULU3

A recent report (Y-W Qian et al., Science 282:1701-1704,1998) described xPlkkl as the activator of Plx1 (the Xenopus Polo kinase). In Xenopus oocytes, the STK Plkk1 can phosphorylate and activate Plx1 STK (the mammalian Polo kinase or PLK). A dominant-negative (kinase-dead) form of xPlkkl prevents Plx1 activation and delays germinal vesicle breakdown. Yet another unidentified kinase is probably responsible for xPlkk1 activation during mitosis.

The homology through the entire length of the xPlkk1 protein with GEK2 suggests that GEK2 might represent the human homologue for xPlkk1. Based on this, GEK2 might be upstream of PLK in mammalian cells. In addition, based on the phage display screen results using the SULU1 coiled-coil2 domain as bait, SULU1 might also interact in vivo with GEK2 and therefore regulate GEK2 (and/or SLK through the coiled-coil domain) activation leading to PLK activation and mitosis.

If such a cascade of events is required for mitosis in mammalian cells, interruption of this signaling cascade at any point might block mitosis and could be beneficial for cancer treatment.

A recently cloned STE20-subfamily kinase, rat TAO1, is most likely the rodent orthologue of human SULU3 (Hutchinson, M. et al. J. Biol. Chem 273:28625-28632, 1998). TAO1 activates MEK3, 4 and 6 in vitro, while in transfected cells it associates and activates only MEK3, resulting in phosphorylation and activation of p38. These results implicate TAO1 (SULU3) in the regulation of the p38 containing stress-responsive MAP kinase pathway.

Human SULU1 is weakly expressed in hematopoietic sources whereas SULU3 is found in B-cells and TH1-restricted T cells. These mammalian SULU STKs display strong homology to the C. elegans SULU kinase. The role that this kinase plays in nematode development is unknown. The strong sequence homology between the catalytic domain of mammalian SULU kinases and other STE20-subfamily kinases such as SOK-1 (human STE20) and KHS2 suggests that the mammalian kinases may participate in the stress-response pathway. The potential coiled-coil domains found at the C-terminus of the SULU kinases may play a role in the regulation of this kinase.

Murine LOK (lymphocyte-oriented kinase) is a constitutively activated STK of approximately 130,000 daltons that is predominantly expressed in spleen, thymus and bone marrow (Kuramochi, S. et al (1997) J. Biol. Chem. 272: 22679-22684) as well as in meiotic testicular and primordial germ cells. The LOK1 gene is located in chromosome 11 of the mouse near the wr locus, a region that is associated with reproductive and neurological defects (Yanagisawa, M. et al (1996) Mol. Reprod. and Dev. 45:411-420). LOK does not activate any of the known MAPK pathways (ERK, JNK and p38) nor the NFκB pathway. The upstream signaling elements of LOK as well as the extracellular stimuli that utilize this kinase to elicit a biological response are also unknown (Kuramochi, S. et al (1997) J. Biol. Chem. 272: 22679-22684).

Human GEK2 is highly related to murine LOK, but based on sequence divergence in the non-catalytic domain, it appears to be a distinct member of this STE20-subfamily. GEK2 may signal through a pathway that remains to be defined. The presence of potential coiled-coil regions at the C-terminus of GEK2 could play a key role in regulating the functions of this kinase.

Human PAK4 and PAK5

The p21 activated protein kinases (PAK) are a closely related subgroup of the STE20 family of serine/threonine kinases. Extensive genetic and biochemical analysis of the budding yeast STE20 has shown the critical role this serine/threonine kinase plays at the juncture of several important intracellular pathways required to appropriately respond to extracellular signals. STE20 links the transcriptional response by mediating the activation of the appropriate downstream MAPK pathway as well as coupling changes in cellular morphology via its control of the actin cytoskeleton.

A hallmark of the PAK subgroup is their small G protein-binding domain (PBD) that confers G protein-dependent activation upon this group of kinases. Via the PBD, PAKs bind to activated small G proteins resulting in the derepression of the PAK's intrinsic kinase activity.

Until recently, there were three known PAK kinases: PAK1, a 68 kD protein whose expression is restricted expression to brain, muscle, and spleen; PAK2 (PAKI, PAK65), a 62 kD protein whose expression is ubiquitous; and PAK3, a 65 kD protein whose expression is restricted to the brain. Similar to STE20, the mammalian PAKs (1,2, and 3) have been shown to respond to extracellular signals (growth factors, mitogens, cytokines and a variety of cellular stresses) (Bagrodia, et al. (1995). J. Biol. Chem. 270: 22731-22737; Zhang, S., et al. (1995). J. Biol. Chem. 270: 23934-23936, Frost, J. et al. (1998) J. Biol. Chem. 273: 28191-28198; Galisteo, M. et al. (1996) J. Biol. Chem. 271: 20997-21000), and are linked to TCR activation (Yablonski, D., et al. (1998) EMBO J. 17: 5647-5657), and heterotrimeric G protein-coupled receptors (Knaus, U. et al. (1995) Science 269: 221-223) The PAKs were originally identified as effectors for members of the Rho family of small G proteins (such as Rac and Cdc42), hence their name, p21-activated kinases (PAK) (Manser et al Nature 367:40-46). The recruitment of the PAKs to the appropriate intracellular location is critical to their function. Attempts to elucidate the role played by PAKs in intracellular signaling and morphological changes is complicated due to the complex interactions by which they can be recruited by such factors as activated small G proteins (rac, cdc42), adaptors (nck) and exchange proteins (PIX, Cool).

The adaptor molecule, Nck, is constitutively bound via its SH3 domain to the proline-rich motif in the N-terminal portion of PAK1. Binding of the Nck-PAK complex to activated growth factor receptors in response to growth factor stimulation provides a mechanism to link growth factor-stimulated and stress-response pathways (Galisteo, M. et al. (1996) J. Biol. Chem. 271:20997-21000).

The PBD found at the N-terminus of PAK1 is responsible for its high-affinity interaction with the GTP-bound forms of Cdc42 and Rac (Burbelo, P. et al. (1995) J. Biol. Chem. 270:29071-29074). The exact mechanism through which the small GTPases activate PAKs may involve, in part, association of the kinase with activated growth factor receptors through guanine nucleotide exchange factors (GEFs). GEFs activate small GTPases by catalyzing the formation of their GTP-bound state, thereby promoting their association with, and activation of, PAKs. The known mammalian PAK kinases, as well as Drosophila and C. elegans PAKs, all conserve an N-terminal extracatalytic motif responsible for a high-affinity interaction with the GEF, PIX. The PAK-Cdc42 interaction and subsequent PAKs occurs as a PIX/PAK complex (Manser, E. et al. (1998) Molecular Cell, 1, 183-192).

PAK signaling stimulated by heterotrimeric G proteins is mediated through the interaction between a short conserved amino acid region located at the C-terminus of PAK1 with the G-protein P-subunit (Leeuw, T. et al. (1998) Nature, 391: 191-195).

A variety of studies have indicated that the human PAKs are involved in mediating the activation of stress-activated protein kinase pathways (JNK and to lesser extent p38). PAKs are also potential mediators in the crosstalk between the pathways regulated by the Rho family of small G proteins and the signaling pathways directly downstream of Ras leading to the activation of the ERK pathway (Bagrodia, et al. (1995). J. Biol. Chem. 270: 22731-22737; Zhang, S., et al. (1995). J. Biol. Chem. 270: 23934-23936; Brown, J., et al. (1996) Curr Biol. 6:598-60596; Frost, J., et al. (1996). Mol. Cell. Biol. 16: 3707-3713).

PAK1 has been implicated in phosphorylating a regulatory site in MEK1 that is necessary for MEK1's ability to interact with Rafl (Frost, et al. (1997) EMBO J. 16:6426-6438). PAK3 has been shown to phosphorylate Rafl on a site that is important for Rafl activity (King, A., et al. (1998). Nature 396: 180-183).

PAKs play an important role in controlling morphological changes in cell shape mediated by the actin cytoskeleton. Such morphological changes are required for cellular functions ranging from cell division and proliferation to cell motility and vesicle transport. PAK activity has been implicated in the localized assembly (leading edge) and disassembly (retracting edge) of focal adhesions necessary for cell motility (Frost J. et al (1998) J. Biol. Chem. 273:28191-28198).

PAK2 may have a role in the morphological changes induced during apoptosis (Membrane and morphological changes in apoptotic cells regulated by caspase-mediated activation of PAK2. (Rudel, T. (1997) Science. 276:1571-4)), and PAK1 may be important in preventing apoptosis (Faure S, et al. (1997) EMBO J. (1997) 16:5550-61). In addition to overcoming mitogen- and anchorage-independent growth, tumor cells need to escape the programmed cell death that accompanies deregulated cell growth. Thus, inhibition of PAKs may be effective in triggering apoptosis in tumors.

A direct requirement for PAKs in the transformation of mammalian cells has been shown for PAK1 and PAK2. Kinase-dead alleles of PAK1 block ras transformation of RAT1 and Schwann cells (Tang, Y., et al. (1997) Mol. Cell. Biol. 17, 4454-4464). Dominant-negative alleles of PAK2 have been shown to interfere with ras-mediated transformation of mammalian cells (Osada, S., (1997) FEBS Lett 404:227-233) Mutations in PAK3 have been implicated in nonsyndromic X-linked mental retardation suggesting a role for PAK3 in cognitive function (Allen, K. et al. (1998) Nat. Genet. 20: 25-30). PAK1 has been implicated in neurite outgrowth in PC12 cells (Daniels, R. et al. (1998) EMBO J. 17: 754-764; Nikolic, M. et al. (1998) Nature 395:194-198).

Finally, PAK-like STKs may also play a role in AIDS pathogenesis since the myristoylated 27 kD membrane-associated HIV Nef gene product directly interacts with and activates these kinases via cdc42 and Rac. The Nef-mediated activation of PAK-like STKs correlates with the induction of high viral titers and the development of AIDS in infected hosts (Cullen, B. R. (1996) Curr. Biol. 6:1557-1559).

Our results show that PAK4 is expressed in thymus, dendrocytes, mast cells, monocytes, as well as in T cells (TH2-restricted cells and MOLT4) and the B cell line RPMI. PAK5 is found in mast cells and in the T cell line MOLT4. These data suggest potential roles for PAK4 and PAKS in the immune system.

PAK4 and PAK5 share with the known PAKs a potential cdc42-binding motif at their N-termini. Both PAK4 and PAK5 display sequence homology in their C-termini to a motif responsible for an interaction between PAK1 and the β-subunit of heterotrimic G-proteins (amino acid residues 665-676 in PAK 4, and amino acid residues 386-398 in PAK5). Consequently, PAK4, and possibly PAK5, could mediate signaling events originating from growth factors as well as from ligands that stimulate G-protein-linked receptors.

PAK4 conserves a leucine (leu 44), that when mutated to a phenylalanine renders the kinase activity of human PAK1 constitutively active, bypassing its cdc42-binding requirement for activation (Brown J. et al (1996) Current Biol. 6:598-605). PAKS contains an isoleucine at the equivalent position. Therefore, the mechanism by which cdc42 potentially activates human PAK1, PAK4, and possibly PAK5, may be very similar.

PAK4 and PAK5 however, lack the PIX-binding motif, and consequently cdc42-activating GEFs other than PIX (for example Dbl and Cool) must be responsible for the activation of these kinases. Alternatively, PAK4 and PAK5 may be activated by another GTPase, such as Racl which uses the Tiaml GEF for its activation to the GTP-bound state.

PAK4 and PAK5 also lack the PxxP (SEQ ID NO: 148) motif responsible for the Nck-PAK1 association. Between the PBD or cdc42-binding N-terminal motifs and the putative GEF-binding regions, PAK4 and PALK5 have long insertions (185 and 123 amino acids for PAK4 and PAKS, respectively) relative to PAK1. This region probably confers different binding characteristics to adaptor molecules and/or GEFs from those exhibited by known mammalian PAKs.

PAKs have been shown to be upstream in pathways leading to activation of both the JNK (Bagrodia, S., et al. (1995) J. Biol. Chem. 270: 22731-22737) and ERK kinase pathways (Brown, J., et al. (1996). Curr Biol. 6:598-605). PAK1 was shown to synergize with ras in activation of the ERK pathway through phosphorylation of MEK1 (Frost, J. et al. (1997). EMBO J. 16:6426-6438). Our data shows that MEK1 serves as an in vitro substrate for PAK4, suggesting a potential role for PAK4 in the activation of the ERK pathway and mitogenesis.

PAK5 may also have a mitogenic role, and be linked to cancer, based on its expression profile (elevated RNA and protein levels in a wide variety of tumor cell lines), its interaction with cdc42 via its PBD, and the ability of a kinase-dead allele (Lys350, 351 Ala) to block ras transformation of NIH3T3 cells. Thus, a screen for small molecule inhibitors of PAK5 kinase activity may yield compounds with therapeutic potential for intervention in cancer derived from a wide variety of tissue types.

PAK4 and PAK5 may also play a role in HIV pathogenesis as potential mediators of Nef signaling, since none of the known PAKs correspond to the PAK-like kinase shown to interact with, and be activated by, the HIV nef protein (Lu, X. et al. (1996) Current Biology 6:1677-1684)

The 3′ untranslated region of PAK4 contains a CA repeat that is prone to undergo expansion. CA dinucleotide repeat instability has been associated with disease (Toren, M.Z. et al (1998) Am. J. Hematol. 57: 148-152), and expansion of such repeat in the 3′ untranslated region of PAK4 could implicate this kinase in as yet unknown pathologies.

Clinical Applications

Human STLK2, STLK3, STLK4, STLK5, STLK6, and STLK7

STLK3, STLK5, STLK6 and STLK7, as well as other homologues of the STLK subfamily of STE20 protein kinases such as STLK4, may play an important role as mediators of the immune response. Thus, they are targets for the development of specific small molecule inhibitors to treat immunological diseases, including, but not limited to, rheumatoid arthritis, chronic inflammatory bowel diseases (e.g. Crohn's disease), chronic inflammatory pelvic disease, multiple sclerosis, asthma, osteoarthritis, psoriasis, atherosclerosis, rhinitis and autoimmunity, as well as in organ transplantation. Other diseases include cardiovascular diseases.

The human STLKs may also play an important role in cell growth regulation. Thus, they are targets for developing small molecule kinase inhibitors for the treatment of cancer and metastases. STLK5 maps to a chromosomal region frequently amplified in a variety of tumors including those from non-small cell lung cancer, breast cancer and peripheral nerve tumors. This suggests that STLK5 could play a role in the development, maintenance, or progression of human tumors.

The potential role of human STLKs 2,3, and 4 in mediating oxidative stress strongly suggests that drugs targeting these kinases could prove useful in the treatment of myocardial infarction, arrhythmia and other cardiomyopathies, stroke, renal failure, oxidative stress-related neurodegenerative disorders such amyotrophic lateral sclerosis, Parkinson's disease and Leigh syndrome, a necrotizing mitochondrial encephalopathy, as well.

Human ZC1, ZC2, ZC3, and ZC4

ZC1 may be a component of the CD28-signaling pathway and therefore important in T cell activation. As such, ZC1 as well as other ZC subfamily kinases, are targets for the development of specific small molecule inhibitors to treat immunological diseases, including, but not limited to, rheumatoid arthritis, chronic inflammatory bowel diseases (e.g. Crohn's disease), chronic inflammatory pelvic disease, multiple sclerosis, asthma, osteoarthritis, psoriasis, atherosclerosis, rhinitis and autoimmunity, as well as organ transplantation. Other diseases include cardiovascular diseases.

ZC1 and ZC2 are also implicated in cell growth regulation. Thus, ZC subfamily kinases are targets for developing small molecule inhibitors for the treatment of cancer and metastases. ZC2 maps to a chromosomal region frequently amplified in a variety of tumors including those from non-small cell lung cancer, small cell lung cancer, and cervical cancer. This suggests that ZC2 could play a role in the development, maintenance, or progression of human tumors.

The role of human ZC1, ZC2, ZC3, and ZC4 in the inflammatory and stress-response pathways, strongly suggests that drugs targeting these kinases could have strong immunosuppressive actions. These drugs can prove valuable for the treatment of rheumatoid arthritis, artherosclerosis, autoimmune disorders and organ transplantation among others. At least one very important class of immunosuppresants, corticosteroids, functions by blocking SAPK activation at an as yet undefined site on this pathway (Swantek, J.L. et al (1997) Mol. Cell. Biol. (1997) 6274-6282). Other immunosuppresive drugs like the pyridinylimidazoles specifically target the p38 kinases (Kumar, S. et al (1997) Biochem. Biophys. Res. Commun. 235: 533-528). Drug targeting of the MAPK and p38 pathways could lead to the development of novel immunosuppresants.

Human SULU and GEK

The potential role of these novel STE20-related protein kinases in the control of mitosis strongly suggests that agents that specifically inhibit these kinases could be useful for cancer and metastases treatment.

The close homology of human STLK5, GEK2, SULU1 and SULU3 to STE20-subfamily kinases involved in the stress and oxidative response pathway strongly suggests that drugs targeting these kinases may also be useful as immunosuppressants as well as to treat ischemic disorders.

Human KHS2

The role of human KHS2 in the inflammatory and stress-response pathways, strongly suggests that drugs targeting this and related kinases could have strong immunosuppressive actions. These drugs can prove valuable for the treatment of rheumatoid arthritis, artherosclerosis, autoimmune disorders and organ transplantation among others. At least one very important class of immunosuppresants, corticosteroids, functions by blocking SAPK activation at an as yet undefined site on this pathway (Swantek, J.L. et al (1997) Mol. Cell. Biol. (1997) 6274-6282). Other immunosuppresive drugs like the pyridinylimidazoles specifically target the p38 kinases (Kumar, S. et al (1997) Biochem. Biophys. Res. Commun. 235: 533-528). Drug targeting of the MAPK and p38 pathways could lead to the development of novel immunosuppressants.

Human PAK Family

PAK5 has a role in cancer based on its expression profile (elevated RNA and protein levels in wide variety of tumor lines), its interaction with Cdc42 via its PBD, and the ability of the kinase-dead allele of PAK5 (Lys350, 351Ala) to block ras transformation of NIH3T3 cells. Thus, a screen for small molecule inhibitors of PAK5 kinase activity may yield compounds with therapeutic potential for intervention in cancers and metastases derived from a wide range of tissue types.

PAK5 maps to a chromosomal region frequently amplified in a variety of tumors including those from non-small cell lung cancer, and small cell lung cancer. These findings suggest that PAK5 could play a role in the development, maintenance, or progression of human tumors and/or metastases.

The role of human PAK4, and PAK5 in the inflammatory and stress-response pathways also strongly suggests that drugs targeting these kinases could have strong immunosuppressive actions. These drugs can prove valuable for the treatment of rheumatoid arthritis, artherosclerosis, autoimmune disorders and organ transplantation among others. At least one very important class of immunosuppresants, corticosteroids, functions by blocking SAPK activation at an as yet undefined site on this pathway (Swantek, J. L. et al (1997) Mol. Cell. Biol. (1997) 6274-6282). Other immunosuppresive drugs like the pyridinylimidazoles specifically target the p38 kinases (Kumar, S. et al (1997) Biochem. Biophys. Res. Commun. 235: 533-528). Drug targeting of the MAPK and p38 pathways could lead to the development of novel immunosuppresants. In addition, drugs targeting PAK4 or PAK5 could prove useful as immunosuppresants as well as in AIDS treatment.

VIII. Transgenic Animals.

A variety of methods are available for the production of transgenic animals associated with this invention. DNA can be injected into the pronucleus of a fertilized egg before fusion of the male and female pronuclei, or injected into the nucleus of an embryonic cell (e.g., the nucleus of a two-cell embryo) following the initiation of cell division (Brinster et al., Proc. Nat. Acad. Sci. USA 82: 4438-4442, 1985). Embryos can be infected with viruses, especially retroviruses, modified to carry inorganic-ion receptor nucleotide sequences of the invention.

Pluripotent stem cells derived from the inner cell mass of the embryo and stabilized in culture can be manipulated in culture to incorporate nucleotide sequences of the invention. A transgenic animal can be produced from such cells through implantation into a blastocyst that is implanted into a foster mother and allowed to come to term. Animals suitable for transgenic experiments can be obtained from standard commercial sources such as Charles River (Wilmington, Mass.), Taconic (Germantown, N.Y.), Harlan Sprague Dawley (Indianapolis, Ind.), etc.

The procedures for manipulation of the rodent embryo and for microinjection of DNA into the pronucleus of the zygote are well known to those of ordinary skill in the art (Hogan et al., supra). Microinjection procedures for fish, amphibian eggs and birds are detailed in Houdebine and Chourrout (Experientia 47: 897-905, 1991). Other procedures for introduction of DNA into tissues of animals are described in U.S. Pat. No. 4,945,050 (Sandford et al., Jul. 30, 1990).

By way of example only, to prepare a transgenic mouse, female mice are induced to superovulate. Females are placed with males, and the mated females are sacrificed by CO₂ asphyxiation or cervical dislocation and embryos are recovered from excised oviducts. Surrounding cumulus cells are removed. Pronuclear embryos are then washed and stored until the time of injection. Randomly cycling adult female mice are paired with vasectomized males. Recipient females are mated at the same time as donor females. Embryos then are transferred surgically. The procedure for generating transgenic rats is similar to that of mice (Hammer et al., Cell 63:1099-1112, 1990).

Methods for the culturing of embryonic stem (ES) cells and the subsequent production of transgenic animals by the introduction of DNA into ES cells using methods such as electroporation, calcium phosphate/DNA precipitation and direct injection also are well known to those of ordinary skill in the art (Teratocarcinomas and Embryonic Stem Cells, A Practical Approach, E. J. Robertson, ed., IRL Press, 1987).

In cases involving random gene integration, a clone containing the sequence(s) of the invention is co-transfected with a gene encoding resistance. Alternatively, the gene encoding neomycin resistance is physically linked to the sequence(s) of the invention. Transfection and isolation of desired clones are carried out by any one of several methods well known to those of ordinary skill in the art (E.J. Robertson, supra).

DNA molecules introduced into ES cells can also be integrated into the chromosome through the process of homologous recombination (Capecchi, Science 244: 1288-1292, 1989). Methods for positive selection of the recombination event (i.e., neo resistance) and dual positive-negative selection (i.e., neo resistance and gancyclovir resistance) and the subsequent identification of the desired clones by PCR have been described by Capecchi, supra and Joyner et al. (Nature 338: 153-156, 1989), the teachings of which are incorporated herein in their entirety including any drawings. The final phase of the procedure is to inject targeted ES cells into blastocysts and to transfer the blastocysts into pseudopregnant females. The resulting chimeric animals are bred and the offspring are analyzed by Southern blotting to identify individuals that carry the transgene. Procedures for the production of non-rodent mammals and other animals have been discussed by others (Houdebine and Chourrout, supra; Pursel et al., Science 244:1281-1288, 1989; and Simms et al., Bio/Technology 6:179-183, 1988).

Thus, the invention provides transgenic, nonhuman mammals containing a transgene encoding a kinase of the invention or a gene effecting the expression of the kinase. Such transgenic nonhuman mammals are particularly useful as an in vivo test system for studying the effects of introduction of a kinase, or regulating the expression of a kinase (i.e., through the introduction of additional genes, antisense nucleic acids, or ribozymes).

A “transgenic animal” is an animal having cells that contain DNA which has been artificially inserted into a cell, which DNA becomes part of the genome of the animal which develops from that cell. Preferred transgenic animals are primates, mice, rats, cows, pigs, horses, goats, sheep, dogs and cats. The transgenic DNA may encode human STE20-related kinases. Native expression in an animal may be reduced by providing an amount of anti-sense RNA or DNA effective to reduce expression of the receptor.

IX. Gene Therapy

STE20-related kinases or their genetic sequences will also be useful in gene therapy (reviewed in Miller, Nature 357:455-460, 1992). Miller states that advances have resulted in practical approaches to human gene therapy that have demonstrated positive initial results. The basic science of gene therapy is described in Mulligan (Science 260:926-931, 1993).

In one preferred embodiment, an expression vector containing STE20-related kinase coding sequence is inserted into cells, the cells are grown in vitro and then infused in large numbers into patients. In another preferred embodiment, a DNA segment containing a promoter of choice (for example a strong promoter) is transferred into cells containing an endogenous gene encoding kinases of the invention in such a manner that the promoter segment enhances expression of the endogenous kinase gene (for example, the promoter segment is transferred to the cell such that it becomes directly linked to the endogenous kinase gene).

The gene therapy may involve the use of an adenovirus containing kinase cDNA targeted to a tumor, systemic kinase increase by implantation of engineered cells, injection with kinase-encoding virus, or injection of naked kinase DNA into appropriate tissues.

Target cell populations may be modified by introducing altered forms of one or more components of the protein complexes in order to modulate the activity of such complexes. For example, by reducing or inhibiting a complex component activity within target cells, an abnormal signal transduction event(s) leading to a condition may be decreased, inhibited, or reversed. Deletion or missense mutants of a component, that retain the ability to interact with other components of the protein complexes but cannot function in signal transduction may be used to inhibit an abnormal, deleterious signal transduction event.

Expression vectors derived from viruses such as retroviruses, vaccinia virus, adenovirus, adeno-associated virus, herpes viruses, several RNA viruses, or bovine papilloma virus, may be used for delivery of nucleotide sequences (e.g., cDNA) encoding recombinant kinase of the invention protein into the targeted cell population (e.g., tumor cells). Methods which are well known to those skilled in the art can be used to construct recombinant viral vectors containing coding sequences (Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y., 1989; Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, N.Y., 1989). Alternatively, recombinant nucleic acid molecules encoding protein sequences can be used as naked DNA or in a reconstituted system e.g., liposomes or other lipid systems for delivery to target cells (e.g., Felgner et al., Nature 337:387-8, 1989). Several other methods for the direct transfer of plasmid DNA into cells exist for use in human gene therapy and involve targeting the DNA to receptors on cells by complexing the plasmid DNA to proteins (Miller, supra).

In its simplest form, gene transfer can be performed by simply injecting minute amounts of DNA into the nucleus of a cell, through a process of microinjection (Capecchi, Cell 22:479-88, 1980). Once recombinant genes are introduced into a cell, they can be recognized by the cell's normal mechanisms for transcription and translation, and a gene product will be expressed. Other methods have also been attempted for introducing DNA into larger numbers of cells. These methods include: transfection, wherein DNA is precipitated with CaPO₄ and taken into cells by pinocytosis (Chen et al., Mol. Cell Biol. 7:2745-52, 1987); electroporation, wherein cells are exposed to large voltage pulses to introduce holes into the membrane (Chu et al., Nucleic Acids Res. 15:1311-26, 1987); lipofection/liposome fusion, wherein DNA is packaged into lipophilic vesicles which fuse with a target cell (Felgner et al., Proc. Natl. Acad. Sci. USA. 84:7413-7417, 1987); and particle bombardment using DNA bound to small projectiles (Yang et al., Proc. Natl. Acad. Sci. 87:9568-9572, 1990). Another method for introducing DNA into cells is to couple the DNA to chemically modified proteins.

It has also been shown that adenovirus proteins are capable of destabilizing endosomes and enhancing the uptake of DNA into cells. The admixture of adenovirus to solutions containing DNA complexes, or the binding of DNA to polylysine covalently attached to adenovirus using protein crosslinking agents substantially improves the uptake and expression of the recombinant gene (Curiel et al., Am. J. Respir. Cell. Mol. Biol., 6:247-52, 1992).

As used herein “gene transfer” means the process of introducing a foreign nucleic acid molecule into a cell. Gene transfer is commonly performed to enable the expression of a particular product encoded by the gene. The product may include a protein, polypeptide, anti-sense DNA or RNA, or enzymatically active RNA. Gene transfer can be performed in cultured cells or by direct administration into animals. Generally gene transfer involves the process of nucleic acid contact with a target cell by non-specific or receptor mediated interactions, uptake of nucleic acid into the cell through the membrane or by endocytosis, and release of nucleic acid into the cytoplasm from the plasma membrane or endosome. Expression may require, in addition, movement of the nucleic acid into the nucleus of the cell and binding to appropriate nuclear factors for transcription.

As used herein “gene therapy” is a form of gene transfer and is included within the definition of gene transfer as used herein and specifically refers to gene transfer to express a therapeutic product from a cell in vivo or in vitro. Gene transfer can be performed ex vivo on cells which are then transplanted into a patient, or can be performed by direct administration of the nucleic acid or nucleic acid-protein complex into the patient.

In another preferred embodiment, a vector having nucleic acid sequences encoding a STE20-related kinase polypeptide is provided in which the nucleic acid sequence is expressed only in specific tissue. Methods of achieving tissue-specific gene expression are set forth in International Publication No. WO 93/09236, filed Nov. 3, 1992 and published May 13, 1993.

In all of the preceding vectors set forth above, a further aspect of the invention is that the nucleic acid sequence contained in the vector may include additions, deletions or modifications to some or all of the sequence of the nucleic acid, as defined above.

In another preferred embodiment, a method of gene replacement is set forth. “Gene replacement” as used herein means supplying a nucleic acid sequence which is capable of being expressed in vivo in an animal and thereby providing or augmenting the function of an endogenous gene which is missing or defective in the animal.

X. Administration of Substances

Methods of determining the dosages of compounds to be administered to a patient and modes of administering compounds to an organism are disclosed in U.S. application Ser. No. 08/702,282, filed Aug. 23, 1996 and International patent publication number WO 96/22976, published Aug. 1, 1996, both of which are incorporated herein by reference in their entirety, including any drawings, figures, or tables. Those skilled in the art will appreciate that such descriptions are applicable to the present invention and can be easily adapted to it.

The proper dosage depends on various factors such as the type of disease being treated, the particular composition being used, and the size and physiological condition of the patient. Therapeutically effective doses for the compounds described herein can be estimated initially from cell culture and animal models. For example, a dose can be formulated in animal models to achieve a circulating concentration range that initially takes into account the ICso as determined in cell culture assays. The animal model data can be used to more accurately determine useful doses in humans.

Plasma half-life and biodistribution of the drug and metabolites in the plasma, tumors, and major organs can be also be determined to facilitate the selection of drugs most appropriate to inhibit a disorder. Such measurements can be carried out. For example, HPLC analysis can be performed on the plasma of animals treated with the drug and the location of radiolabeled compounds can be determined using detection methods such as X-ray, CAT scan, and MRI. Compounds that show potent inhibitory activity in the screening assays, but have poor pharmacokinetic characteristics, can be optimized by altering the chemical structure and retesting. In this regard, compounds displaying good pharmacokinetic characteristics can be used as a model.

Toxicity studies can also be carried out by measuring the blood cell composition. For example, toxicity studies can be carried out in a suitable animal model as follows: 1) the compound is administered to mice (an untreated control mouse should also be used); 2) blood samples are periodically obtained via the tail vein from one mouse in each treatment group; and 3) the samples are analyzed for red and white blood cell counts, blood cell composition, and the percent of lymphocytes versus polymorphonuclear cells. A comparison of results for each dosing regime with the controls indicates if toxicity is present.

At the termination of each toxicity study, further studies can be carried out by sacrificing the animals (preferably, in accordance with the American Veterinary Medical Association guidelines Report of the American Veterinary Medical Assoc. Panel on Euthanasia, Journal of American Veterinary Medical Assoc., 202:229-249, 1993). Representative animals from each treatment group can then be examined by gross necropsy for immediate evidence of metastasis, unusual illness, or toxicity. Gross abnormalities in tissue are noted, and tissues are examined histologically. Compounds causing a reduction in body weight or blood components are less preferred, as are compounds having an adverse effect on major organs. In general, the greater the adverse effect the less preferred the compound.

For the treatment of cancers the expected daily dose of a hydrophobic pharmaceutical agent is between 1 to 500 mg/day, preferably 1 to 250 mg/day, and most preferably 1 to 50 mg/day. Drugs can be delivered less frequently provided plasma levels of the active moiety are sufficient to maintain therapeutic effectiveness.

Plasma levels should reflect the potency of the drug. Generally, the more potent the compound the lower the plasma levels necessary to achieve efficacy.

EXAMPLES

The examples below are not limiting and are merely representative of various aspects and features of the present invention. The examples below demonstrate the isolation and characterization of the STE20-related kinases of the invention.

Example 1 Isolation of cDNAs Encoding Mammalian STE20-related Protein Kinases

Materials and Methods

Identification of Novel Clones

Total RNAs were isolated using the Guanidine Salts/Phenol extraction protocol of Chomczynski and Sacchi (P. Chomczynski and N. Sacchi, Anal. Biochem. 162, 156 (1987)) from primary human tumors, normal and tumor cell lines, normal human tissues, and sorted human hematopoietic cells. These RNAs were used to generate single-stranded cDNA using the Superscript Preamplification System (GIBCO BRL, Gaithersburg, Md.; Gerard, G F et al. (1989), FOCUS 11, 66) under conditions recommended by the manufacturer. A typical reaction used 10 μg total RNA with 1.5 μg oligo(dT)₁₂₋₁₈ in a reaction volume of 60 μL. The product was treated with RNaseH and diluted to 100 μL with H₂O. For subsequent PCR amplification, 1-4 μL of this sscDNA was used in each reaction.

Degenerate oligonucleotides were synthesized on an Applied Biosystems 3948 DNA synthesizer using established phosphoramidite chemistry, precipitated with ethanol and used unpurified for PCR. The sequence of some of the degenerate oligonucleotide primers and the amino acid motif they encode is as follows:

TRK1 5′-CTGAATTCGGNGCNTTYGGNAARGT-3′ (SEQ ID NO:32)    GAFGKV (sense) (SEQ ID NO:37) TRK4 5′-GCTGGATCCYTCNGGNGGCATCCA-3′ (SEQ ID NO:33)    WMPPE (antisense) (SEQ ID NO:38) ROS1 5′-GCNTTYGGNGARGTNTAYGARGG-3′ (SEQ ID NO:34)    AFGEVYEG (sense) (SEQ ID NO:39) CCK4b 5′-GCTGGATCCYTCNGGNSWCATCCA-3′ (SEQ ID NO:35)    WMSPE (antisense) (SEQ ID NO:40) CCK4c 5′-GAGTTYGGNGARGTNTTYYTNGC-3′ (SEQ ID NO:36)    EFGEVYEG (sense) (SEQ ID NO:41)

These primers were derived from the sense and antisense strands of conserved motifs within the catalytic domain of several protein kinases. Degenerate nucleotide residue designations are: N=A, C, G, or T; R=A or G; Y=C or T; H=A, C or T not G; D=A, G or T not C; S=C or G; and W=A or T.

PCR reactions were performed using degenerate primers applied to multiple single-stranded cDNAs. The primers were added at a final concentration of 5 μM each to a mixture containing 10 mM TrisHCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl₂, 200 μM each deoxynucleoside triphosphate, 0.001% gelatin, 1.5 U AmpliTaq DNA Polymerase (Perkin-Elmer/Cetus), and 1-4 μL cDNA. Following 3 min denaturation at 95° C., the cycling conditions were 94° C. for 30 s, 50° C. for 1 min, and 72° C. for 1 min 45 s for 35 cycles. PCR fragments migrating between 300-350 bp were isolated from 2% agarose gels using the GeneClean Kit (Bio101), and T-A cloned into the pCR11 vector (Invitrogen Corp. U.S.A.) according to the manufacturer's protocol.

Colonies were selected for mini plasmid DNA-preparations using Qiagen columns and the plasmid DNA was sequenced using a cycle sequencing dye-terminator kit with AmpliTaq DNA Polymerase, FS (ABI, Foster City, Calif.). Sequencing reaction products were run on an ABI Prism 377 DNA Sequencer, and analyzed using the BLAST alignment algorithm (Altschul, S. F. et al., J.Mol.Biol. 215: 403-10).

Additional PCR strategies were employed to connect various PCR fragments or ESTs using exact or near exact oligonucleotide primers as detailed in the results section for each cDNA. PCR conditions were as described above except the annealing temperatures were calculated for each oligo pair using the formula: Tm=4(G+C)+2(A+T).

Isolation of cDNA Clones:

Human cDNA libraries were probed with PCR or EST fragments corresponding to STE20-related genes. Probes were ³²P-labeled by random priming and used at 2×10⁶ cpm/mL following standard techniques for library screening. Pre-hybridization (3 h) and hybridization (overnight) were conducted at 42° C. in 5× SSC, 5× Denhart's solution, 2.5% dextran sulfate, 50 mM Na₂PO₄/NaHPO₄, pH 7.0, 50% formamide with 100 mg/mL denatured salmon sperm DNA. Stringent washes were performed at 65° C. in 0.1× SSC and 0.1% SDS. DNA sequencing was carried out on both strands using a cycle sequencing dye-terminator kit with AmpliTaq DNA Polymerase, FS (ABI, Foster City, Calif.). Sequencing reaction products were run on an ABI Prism 377 DNA Sequencer.

Makegene Bioinformatics EST assembler

The EST reports were downloaded from ncbi (National Institute for Biotechnology Information). After uncompressing the files, the program ‘report2est’ was scripted to extract the following information: 1) EST names, 2) GenBank Accession numbers, 3) GenBank gi numbers, 4) Clone Id numbers, 5) the nucleotide sequences of the ESTs 6) the organism, 7) the library name, 8) the name of the lab, and 9) the institution. The output of ‘report2est’ is a file in FASTA format with all of the information listed above in the first line of each entry except the sequence, which is listed in the second line of each entry. The resulting file is formatted for BLAST using ‘pressdb’ (available as part of the ncbi tool kit).

To build a gene or part of a gene from ESTs, the program ‘makegene’ was developed. Input to this program is a query sequence and the organism/species for which a gene is to be built. An initial search of the formatted EST database described above is performed using BLAST (blastn). Any results that contain warnings, such as polyA tails or other repeat elements, are eliminated from future queries. The program ‘blast_parse_reports’ was developed to extract the FASTA header line from the search results and the output is then filtered to extract only FASTA header lines for the desired species.

The initial results, having been filtered for warnings and species, go into a loop in which searches against the database are repeated until no new ESTs are found. The loop consists of the following steps: 1) when possible the names of both ends of the ESTs are extracted from the database by searching using the ‘Clone Id’ field or the part of the ‘EST name’ field before the .r or s postscript, 2) any ESTs that have been used as queries in previous loops are removed from the current query by the program ‘subtract’, 3) the resulting list of ESTs is used to extract the sequences from the database by the program batch_parse_fasta, 4) BLAST is run against the database using each sequence, 5) the output files from BLAST containing warnings are removed, 6) the results are filtered by species, and 7) the loop is reentered if there were new ESTs found in the previous pass through the loop.

The ESTs chosen by ‘makegene’ are used as input for the program ‘mpd2 cluster’ (Hide, W., Burke, J, and Davison, D. U. of Houston, unpublished) which clusters overlapping sequences. The programs ‘contig’ (Kerlavage, T., TIGR, unpublished), ‘gde2mult’ and ‘gde2sing’ (Smith, S. W., et al., CABIOS 10, 671-675 (1994)), are used to make an alignment and consensus sequence of the overlapping ESTs.

Results

cDNA Cloning and Characterization of STLK2

The human STLK2 cDNA sequence is composed of two overlapping EST fragments, AA191319 and W16504, that were identified using a Smith-Waterman search of the EST database with STLK1 (MST3 GB:AF024636) as a query. The complete sequence of both clones was determined and used to generate the full-length human STL2 sequence.

EST clone AA191319 contains a 1327 bp insert and an ORF of 1146 bp (382 amino acids). EST clone W16504 contains a 2474 bp insert (not including the poly-A tail) and an ORF of 687 bp (382 amino acids).

The full-length human STLK2 cDNA (SEQ ID NO. 1) is 3268 bp long. AA191319 spans positions 1-1327 and W16504 positions 743-3216. The overlap between these two clones exhibits 100% sequence identity. The human STLK2 cDNA constains a 1248 bp ORF flanked by a 181 bp 5′ UTR (1-181) and a 1784 bp 3′ UTR (1433-3216) that is followed by a 52 nucleotide polyadenylated region. A polyadenylation signal (AATAAA) is found at positions 3193-3198. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for STLK2. Furthermore, human STLK2, and the related SOK-1 and MST3 proteins, conserve the amino acid sequence immediately following this presumed initiating methionine.

Several EST fragments span the complete STLK2 sequence with AA191319 at the 5′ end and W16504 at the 3′ end.

All searches against the public nucleic acid database (NRN) and protein database (NRP) were conducted using the Smith-Waterman gap alignment program ((Smith, T F and Waterman, M S (1981) J. Mol. Biol, 147, 195-197).) with the PAM100 matrix and gap open and extension penalties of 14:1, respectively.

cDNA Cloning and Characterization of STLK3

A mammalian STLK3 clone, 135-31-19, was first identified from a PCR screen with the degenerate oligos, TRK1 and TRK4, applied to a sscDNA generated from adult rat brain substantia nigra. Sequence analysis of the 457 bp insert indicated that it represented a novel member of the STE20-subfamily of STKs.

A Smith-Waterman search of the EST database with the rat STLK3 fragment and human STLK1 (MST3 GB:AF024636) as queries identified several overlapping ESTs spanning most of the human STLK3 cDNA sequence. A Makegene analysis generated a 3037 bp contig from approximately 44 EST sequences. Since the 3′ ESTs were not commercially available, a pair of primers (5′-CACAGAAACGGTCAGATTCAC-3′(SEQ ID NO: 42) and 5t-GATCAGGGTGACATCAAGGGAC-3′(SEQ ID NO: 43)) were derived from this region to generate PCR clone 3R21-20-6 from human fetal liver sscDNA. This clone and EST AA278967 were fully sequenced to generate the full-length STLK2 cDNA sequence.

AA278967 is a 837 bp EST isolated by the IMAGE consortium from cDNA made from CD20+/IgD-germinal center B cells sorted from human tonsillar cells.

PCR clone 3R21-20-6 was isolated from human fetal sscDNA and contains a 1116 bp insert, including a 1086 bp ORF encoding the 362 C-terminal amino acids of STLK3.

The full-length human STLK3 cDNA (SEQ ID NO. 2) is 3030 bp long. AA278967 spans positions 1-814 and 3R21-20-6 spans positions 464-1579. The overlap between these two clones exhibits 100% sequence identity. The remaining 1452 bp of 3′ UTR is derived from an assembly of multiple unconfirmed EST fragments.

The near full-length human STLK3 cDNA (SEQ ID NO.2) is 3030 bp long and consists of a 1548 bp ORF flanked by a 1476 bp 3′ UTR (1550-3025) and a 5 nucleotide polyadenylated region. A polyadenylation signal (AATAAA) begins at position 3004. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. Six copies of a “GGCCCC” repeat were observed in positions 21-67. Five independent ESTs (AA150838, AA286879, AA251679, AA252004, AA278967) showed the same repeat, suggesting that this sequence may be an integral region of the human STLK3 gene. Stronger evidence for this being the case is provided by the sequence of the murine orthologue of STLK3 represented by a 876 bp EST W20737.

Multiple EST fragments span the complete STLK3 sequence with AA278967 at the 5′ end and AA628477 and others at the 3′ end.

cDNA Cloning and Characterization of STLK4

The human STLK4 cDNA sequence is composed of two overlapping EST fragments, AA297759 and AA100484, that were identified using a Smith-Waterman search of the EST database with STLK1 (MST3 GB:AF024636) as a query. The complete sequence of both clones was determined and used to generate the near full-length human STLK4 sequence.

AA100484 is an IMAGE consortium cDNA clone isolated from the T-84 colonic epithelium cell line. It has an insert of 3694 bp and a coding region of 1146 bp (382 amino acids). A Smith-Waterman sequence alignment against the NRN database showed this EST to be 71.4% identical to the human STE20-like kinase (GB:X99325).

W16504 is an IMAGE consortium clone isolated from a human fetal heart cDNA library. It has an insert length of 2474 bp (not including the poly-A tail) and a coding region of 687 bp (229 amino acids). A Smith-Waterman sequence alignment of W16504 against the NRN database showed this EST to be 69.2% identical to the human STE20-like kinase (GB:X99325).

The full-length human STLK2 cDNA (SEQ ID NO. 1) is 3268 bp long. AA191319 spans positions 1-1327, and W16504 positions 743-3216. The overlap between these two clones is 585 bp long with 100% sequence identity.

AA100484 is an IMAGE consortium cDNA clone isolated from the T-84 colonic epithelium cell line. AA100484 covers the bulk of Human STLK4 with its 3694 bp, which spans positions 146-3839 of SEQ ID NO:3. A second EST, AA297759, isolated from a Jurkat T cell cDNA library, spans positions 1-271 of the human STLK4 contig. The two ESTs overlap over a 126 bp stretch that has only one nucleotide discrepancy at position 149 (G in AA297759 and T in AA100484). A T at this position was chosen for the SEQ ID NO:3 based on sequence data generated from A100484. The 5′ 145 bp of human STLK4 contains three sequencing ambiguities (N's in SEQ ID NO:3) arising from sequence errors in the GenBank entry for AA297759. Three amino acid sequence ambiguities in the N-terminus of human STLK4 are present also in SEQ ID NO:7 as a consequence of the sequence inaccuracies from the EST entry.

The coding region of human STLK4 is 1242 bp long (2-1243), capable of encoding a 414 amino acid polypeptide, and is followed by a 2596 nucleotide 3′ UTR (1244-3839). Human STLK4 ends in a polyadenylated stretch that has 18 adenines (3840-3857). A polyadenylation signal (AATAAA) is found between positions 3822-3827. Targeted-PCR cloning identified one rat orthologue of human STLK4, clone 135-31-19. In addition, one murine orthologue of human STLK4 was recognized in the EST database as AA117483. None of these orthologues add additional N-terminal sequence to the human STLK4.

The near full-length human STLK4 cDNA (SEQ ID NO.3) is 3857 bp long and consists of a 1242 bp ORF flanked by a 2596 bp 3′ UTR (1244-3839) and an 18 nucleotide polyadenylated region. Polyadenylation signals (AATAAA) begin at positions 2181 and 3822. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine. A near full-length murine STLK4 cDNA is represented in the 1773 bp EST AA117438. It extends an additional 21 nucleotides 5′ of the human STLK4 consensus, but since its coding region is open throughout the 5′ extent of the sequence, this is also probably a partial cDNA clone lacking the N-terminal start methionine.

Several EST fragments span the complete STLK3 sequence with AA297759 at the 5′ end and AA100484 and others at the 3′ end.

cDNA Cloning and Characterization of STLK5

The human STLK5 cDNA sequence is composed of four overlapping sequences, AI418298, 2R96-13-1, 3R25-45-3 and R46685. A human STLK5 clone, F07734, was first identified using a Smith-Waterman search of the EST database with SPS_sc (U33057) as a query.

AI418298 is an IMAGE consortium cDNA clone with an 895 bp insert.

PCR clone 2R96-13-1 was isolated from human brain sscDNA using primers 5′-CTCATCTGTACACACTTCATGG(SEQ ID NO:44) and 5′-GATTCCCACACTGTAGATGTC(SEQ ID NO:45) derived from F07734.2R96-13-1 contains a 330 bp insert and an ORF of 330 bp (110 amino acids).

EST clone R46685 was identified using a Smith-Waterman search of the EST database with the C-terminus of SPS_sc (GB:U33057) as query. Sequence analysis of the 1047 bp insert identified this EST to contain an ORF of 285 bp (95 amino acids) encoding the C-terminus of human STLK5.

PCR clone 3R25-45-3 was isolated from human fetal brain sscDNA using primers 5′-GGCCCTCGACTACATCCACCACAT(SEQ ID NO:46) and 5′-CAACGAAACTAACACAGCATAAGG(SEQ ID NO:47) derived from 2R96-13-1 and R46685, respectively. 3R25-45-3 contains a 330 bp insert and an ORF of 750 bp (250 amino acids).

The full-length human STLK5 cDNA (SEQ ID NO:96) is 2110 bp long and consists of a 1119 bp ORF flanked by a 229 bp 5′ UTR and a 762 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (supra) for an initiating methionine, and is believed to be the translational start site for STLK5.

Several EST fragments span the complete STLK5 sequence with AA297059 and F07734 at the 5′ end and R46686 and F03423 and others at the 3′ end.

STLK5 displays a 100% match over a 41 bp stretch (position 2-42, SEQ ID NO. 97) to a human CpG island repeat (Z61277).

cDNA Cloning and Characterization of STLK6

Human STLK6 was first identified in the translated EST database (AA219667) as a novel serine threonine kinase.

The partial human STLK6 cDNA (SEQ ID NO:98) is 2,001 bp long and consists of a 1,254 bp ORF flanked by a 75 bp 5′ UTR and a 673 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (Kozak, M., Nucleic Acids Res. 15, 8125-8148 (1987)) for an initiating methionine, and is believed to be the translational start site for STLK6.

At the time of filing, inventors believe that STLK6 does not have any significant match in the nucleic acid database.

cDNA Cloning and Characterization of STLK7

Human STLK7 was first identified in the translated EST database (AA988954) as a novel serine threonine kinase. The original clone was not available through public sources, so a PCR fragment amplified from the sequence of AA988954 yielded 5R54-21-2.

The partial human STLK7 cDNA (SEQ ID NO:100) is 311 bp long and consists of a 309 bp ORF. Since the coding region is open throughout the 5′ and 3′ extent of this sequence, this appears to be a partial cDNA clone lacking the N-terminal start methionine and C-terminal stop codon.

STLK7 shares 80% sequence identity to human SPAK (AF099989) over a 167 bp region and 50% nucleotide sequence identity to SLTK7 (SEQ ID NO. 101) over 391 nucleotides.

cDNA Cloning and Characterization of ZC1

The human ZC1 cDNA sequence is composed of two overlapping PCR clones, 3R25-24-2 and R65-12-2.

A human ZC1 clone, 125-33-5, was first identified from a PCR screen with degenerate oligos, TRK1 and TRK4, applied to sscDNA generated from human small airway epithelial cells (Clontech). Sequence analysis of the 503 bp insert identified a 501 bp ORF (167 amino acids) with the potential to encode a novel human STK related to the C. elegans ZC504.4 gene product.

PCR clone 3R25-24-2 was isolated from human SNB19 glioblastoma sscDNA using primers 5′-ATGGCGAACGACTCTCCCGCGAA(SEQ ID NO:48) and 5′-ACACCAAAATCAACAAGTTTCACCTC(SEQ ID NO:49) derived from the N-terminus of a murine orthologue ofZCl (NIK, GB:U88984) and the original human ZC1 clone 125-33-5, respectively. 3R25-24-2 contains a 527 bp insert and an ORF of519 bp (173 amino acids).

PCR clone R65-12-2 was isolated as follows: A Smith-Waterman search of the EST database with the C. elegans ZC504.4 gene (GB: Z50029) as a query identified a human EST (W81656) whose ORF is related to the C. elegans gene and terminates in an identical residue (Trp). A primer was designed 3′ to this stop codon (5′-AGTTACAAGGAATTCCAAGTTCT(SEQ ID NO:50)) and used in a PCR reaction with a primer derived from the original human ZC1 clone 125-33-5 (5′-ATGAAGAGGAAGAAATCAAACTG(SEQ ID NO:51)) using sscDNA from human SNB19 glioblastoma as a template. PCR clone R65-12-2 was identified and was found to contain a 3611 bp insert with a 3534 bp ORF encoding the C-terminal portion of human ZC1 (1178 amino acids).

The full-length human ZC1 cDNA (SEQ ID NO. 9) is 3798 bp long. Clone 3R25-24-2 spans positions 1-527, and clone R65-12-2 spans positions 188-3798. The overlap between these two clones exhibits 100% sequence identity. The human ZC1 contains a 3717 bp ORF (17-3723) flanked by a 6 bp 5′ UTR and a 75 bp (3724-3798) 3′ UTR. No polyadenylation signal (AATAAA) or polyadenylated region are present in the 3′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human ZC1.

Multiple EST fragments (W81656) match the 3′ end of the human ZC1 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of ZC2

The human ZC2 cDNA sequence is composed of four overlapping PCR clones, G75-31-17, R65-24-6, 2R28-8-1, and R99-6-10.

A human ZC2 clone, G75-31-17, was first identified from a PCR screen with degenerate oligos, ROSi (5′-GCNTTYGGNGARGTNTAYGARGG(SEQ ID NO:34)) and CCK4b (5′-GCTGGATCCYTCNGGNSWCATCCA(SEQ ID NO:35)), applied to sscDNA generated from the human HLT383 primary non-small cell lung cancer tissue. Sequence analysis of the 492 bp insert identified a 492 ORF (164 amino acids) with the potential to encode a novel human STK related to the C. elegans ZC504.4 gene product.

PCR clone R99-6-10 was isolated as follows: A Smith-Waterman search of the EST database with C. elegans ZC504.4 gene (GB: Z50029) as a query identified two overlapping human EST fragments (AA115844 and R51245) whose ORFs were related to the C. elegans gene and terminate in an identical residue (Trp). A primer was designed 3′ to the stop codon found in R51245 (5′-AGATGGACTGTACTGGGAGG(SEQ ID NO:52)) and used in a PCR reaction with a primer derived from AA115844 (5′-ACTTTGTGCAGCTCTGTGGG(SEQ ID NO:53)) using human fetal brain sscDNA as a template. PCR clone R99-6-10 was identified and was found to contain a 1095 bp insert with a 930 bp ORF encoding the C-terminal portion of human ZC2 (310 amino acids).

PCR clone R65-24-6 was isolated from human HT29 colon cancer cell line sscDNA using primers 5′-AAGGTTATGGATGTCACAGGG(SBQ ID NO:54) and 5′-AGATGGACTGTACTGGGAGG(SEQ ID NO:52) derived from G75-31-17 and R51245, respectively. The 3′ primer used in this PCR reaction misprimed between positions 1634-1653 of this gene leading to the formation of a truncated product. R65-24-6 contains a 1593 bp insert and an ORF of 1593 bp (531 amino acids).

PCR clone 2R28-8-1 was isolated from human colon cancer cell line 14T29 sscDNA using primers 5′-CTCACAAGGTTGCCAACAGG(SEQ ID NO:55) and 5′-AGTCCCCACCAGAAGGTTTAC(SEQ ID NO:56) derived from R65-24-6 and R99-6-1O, respectively. 2R28-8-1 contains a 1538 bp insert and an ORF of 1536 bp (512 amino acids).

The partial human ZC2 cDNA (SEQ ID NO. 10) is 4055 bp long. Clone G75-31-17 spans positions 1-492, clone R65-24-6 spans positions 58-1650, clone 2R28-8-1 spans positions 1466-3003 and clone R99-6-10 spans positions 2961-4055. The overlaping regions between these clones exhibit 100% sequence identity except for a single guanine (G75-31-17) to adenosine (R65-24-6) mismatch at position 280 resulting in a Glu to Lys change. Based on the presence of an acidic residue in this position in human ZC1 and ZC3 and C. elegans ZC504.4, the sequence encoding the Glu is probably correct. The human ZC2 gene contains a 3891 bp ORF (1-3891) flanked by 164 bp (3892-4055) 3′ UTR. No polyadenylation signal (AATAAA) or polyadenylated region is present in the 3′UTR.

Multiple EST fragments (R51245) match the 3′ end of the human ZC2 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of ZC3

The human ZC3 cDNA sequence is composed of four overlapping PCR clones, G75-30-30, 3R33-5-3, 3R19-17-6, and R99-43-11.

A human ZC3 clone, G75-30-30, was first identified from a PCR screen with degenerate oligos, ROS1 and CCK4b, applied to sscDNA generated from a human HLT370 primary non-small cell lung cancer tissue. Sequence analysis of the 492 bp insert identified a 492 ORF (164 amino acids) with the potential to encode a novel human STK related to the C. elegans ZC504.4 gene product.

PCR clone R99-43-11 was isolated as follows: A Smith-Waterman search of the EST database with the C. elegans ZC504.4 gene (GB: Z50029) as a query identified a human EST (R54563) whose ORF is related to the C. elegans gene and terminates in an identical residue (Trp). A primer was designed 3′ to the stop codon found in R54563 (5′-TCAGGGGTCAGAGGTCACG(SEQ ID NO:57)) and used in a PCR reaction with a primer derived from the 5′ end of R54563 (5′-CCCAAACCCTACCACAAATTC(SEQ ID NO:58)) using sscDNA from human fetal brain as a template. PCR clone R99-43-11 was identified and was found to contain a 719 bp insert with a 564 bp ORF encoding the C-terminal portion of human ZC3 (188 amino acids).

PCR clone 3R19-17-6 was isolated from human A549 lung cancer cell line sscDNA using primers 5′-CCCCCGGGAAACGATGACCA(SEQ ID NO:59) and 5′-AGCCGCTGCCCCTCCTCTACTGT(SEQ ID NO:60) derived from G75-30-30 and R99-43-11, respectively. The 3′ primer used in this PCR reaction misprimed leading to the formation of a truncated product. 3R19-17-6 contains a 1172 bp insert and an ORF of 1170 bp (390 amino acids).

PCR clone 3R33-5-3 was isolated from human A549 lung cancer cell line sscDNA using primers 5′-ACCGCAACATCGCCACCTACTAC(SEQ ID NO:61) and 5′-CTCGACGTCGTGGACCACC(SEQ ID NO:62) derived from G75-30-30 and 3R19-17-6, respectively. 3R33-5-3 contains a 2465 bp insert and an ORF of 2463 bp (821 amino acids).

The full-length human ZC3 cDNA (SEQ ID NO. 11) is 4133 bp long. Clone G75-30-30 spans positions 1-483, clone 3R33-5-3 spans positions 134-2598, clone 3R19-17-6 spans positions 2356-3512 and clone R99-43-11 spans positions 3415-4133. The overlaps between these clones exhibit 100% sequence identity. The human ZC3 gene contains a 3978 bp ORF (1-3978) flanked by a 152 bp 3′ UTR (3979-4133). No polyadenylation signal (AATAAA) or polyadenylated region is present in the 3′UTR.

Multiple EST fragments (R54563) match the 3′end of the human ZC3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of ZC4

The human ZC4 cDNA sequence, represented by PCR fragment 3R25-27-1, was first identified in the human genomic cosmid 82J11 (GB:Z833850) containing exon sequences that displayed strong homology to the ZC504.4 C. elegans gene.

PCR clone 3R25-27-1 was isolated from human fetal liver sscDNA and primers 5′-CAATGTTAACCCACTCTATGTCTC(SEQ ID NO:63) and 5′-AGTTTGCCGATGTTTTTCTTTTC(SEQ ID NO:64) derived from a potential ORF (positions 25729-25852) from the 82J11 cosmid and from an EST (R98571) encoding the C-terminus of the human ZC4 gene, respectively.

The partial human ZC4 cDNA (SEQ ID NO.12) is 1459 bp long and consists of a 1047 bp ORF (2-1048) flanked by a 411 bp (1049-1459) 3′UTR region. No polyadenylation signal (AATAAA) or polyadenylated region is present in the 3′UTR.

The N-terminal coding sequence for ZC4_h was extended by building a contiguous DNA sequence of 233,137 bp containing Z83850 and four other sequences: cU84B10 and cU230B10 (from the Sanger Human Genome Sequencing Project, and Z97356 and Z69734 (available from the National Institute for Biotechnology Information,). The position of each sequence in the contig is represented in the table below.

Accession Length Start End cU84B10 43273   0  43273 Z97356 21848 43171  65018 Z69734 37077 63073 100149 cU230B10 11841 88416 100256 Z83850 132981  100156  233137

Sequences in ZC4 genomic contig.

The 233,137 bp contig was analyzed for exons using the programs FGENES 1.5 and FGENESH, human gene structure prediction software available from The Sanger Centre.

The resulting human ZC4 coding sequence (SEQ ID NO:104) is 3,681 bp long (excluding the stop codon) and encodes for a STE20 kinase of 1227 amino acids.

cDNA Cloning and Characterization of KHS2

The human KHS2 cDNA sequence is composed of four overlapping clones, 3R25-51-2, 3R16-34-2, 3R16-31-2, and T79916.

A human KHS2 clone, AA250855, was first identified using a Smith-Waterman search of the EST database with KHS1 (GB:U77129) as a query. Sequence analysis of the 1112 bp insert identified a 618 bp ORF (206 amino acids) with the potential to encode a novel STK related to the human KHS1 gene product. Using AA250855 as a query, a second EST (AA446022) was found whose sequence was shown to contain the initiator methionine for human KHS2 based on a comparison with KHS1.

PCR clone 3R25-51-2 was isolated from human testicular cancer sscDNA using primers 5′-CCGCCATGAACCCCGGCTT(SEQ ID NO:65) and 5′-CGATTGCCAAAGACCGTGTCA(SEQ ID NO:66) derived from AA446022 and AA250855, respectively. 3R25-51-2 contains an 850 bp insert and an ORF of 849 bp (283 amino acids).

EST clone, T79916, was identified using a Smith-Waterman search of the EST database with the C-terminus of KHS1 (GB:U77129) as a query. Sequence analysis of the 2107 bp insert identified this EST to contain an ORF of 345 bp (115 amino acids disrupted by a single stop codon) encoding the C-terminus of human KHS2, followed by 1762 bp 3′UTR.

PCR clone 3R16-34-2 was isolated from human testis sscDNA using primers 5′-AGAAGTTGCAGCTGTTGAGAGGA(SEQ ID NO:67) and 5′-TATGGCCCGTGTAAGGATTTC(SEQ ID NO:68) derived from AA250885 and T79916, respectively. 3R16-34-2 contains an 1516 bp insert and an ORF of 1128 bp (376 amino acids).

PCR clone 3R16-31-2 was isolated from normal human colon sscDNA using primers 5′-GTGCCAGAAGTGTTGTGTTGTAA(SEQ ID NO:69) and 5′-TATGGCCCGTGTAAGGATTTC(SEQ ID NO:68) derived from EST T79916.3R16-31-2 contains a 728 bp insert and an ORF of 669 bp (223 amino acids). This clone lacked the stop codon present within EST T79916 (postion 2662 in the KHS2 sequence).

The full-length human KHS2 cDNA (SEQ ID NO.17) is 4023 bp long. Clone 3R25-51-2 spans positions 1-855, clone AA250885 spans positions 336-923, clone 3R16-34-2 spans positions 545-2061, and clone T79916 spans positions 1917-4023. The overlaping regions between these clones exhibit 100% sequence identity, except for 4 nucleotide differences, two of which are silent, a third corrects the internal stop codon at position 2662, and the fourth at position 247 (T to C change) results in a Pro to Leu change. The human KHS2 cDNA contains a 2682 bp ORF (6-2687) flanked by a 5 bp (1-5) 5′UTR and a 1336 bp (2688-4023) 3′ UTR. A potential polyadenylation signal (AATAAA) is found at positions 4008-4013. No polyadenylated region is present in the 3′UTR. The sequence flanking the first ATG is in a poor context for translational initiation, however, a 134 bp 5′UTR sequence from EST AA446022 did not reveal any additional ATG's and displayed two in-frame stop codons 5′ to the putative start ATG for human KHS2.

Multiple EST fragments match the 5′end (AA446022) as well as the 3′ end (R37625) of the human KHS2 gene.

cDNA Cloning and Characterization of SULU1

The human SULU1 cDNA sequence is composed of three overlapping clones, N40091, 2R90-1-1 and R90907.

A human SULU1 clone, N40091, was first identified using a Smith-Waterman search of the EST database with the C. elegans SULU gene (GB: U32275) as a query. Sequence analysis of the 1321 bp insert identified a 906 bp ORF (302 amino acids) with the potential to encode a novel human STK related to the C. elegans SULU gene product.

EST clone R90907 was first identified using a Smith-Waterman search of the EST database with the 3′ end of the C. elegans SULU gene (GB: U32275) as a query. Sequence analysis of the 1647 bp insert identified a 578 bp ORF (192 amino acids) with the potential to encode the C-terminus of the human SULU1 gene product.

PCR clone 2R90-1-l was isolated from human HT29 colon cancer cell sscDNA using primers 5′-TATTGAATTGGCGGAACGGAAG(SEQ ID NO:70) and 5′-TTGTTTTGTGCTCATTCTTTGGAG(SEQ ID NO:71) derived from N40091 and R90907, respectively. 2R90-1-1 contains a 1625 bp insert and an ORF of 1623 bp (541 amino acids).

The full-length human SULU1 cDNA (SEQ ID NO.19) is 4177 bp long Clone N40091 spans positions 1-1321, clone 2R90-1-1 spans positions 1048-2671, and clone R90907 spans positions 2531-4177. The overlaping regions between these clones exhibit 100% sequence identity. The human SULU1 cDNA contains a 2694 bp ORF (415-3108) flanked by a 414 bp (1-414) 5′UTR and a 1069 bp (3109-4177) 3′ UTR followed by a 19 nucleotide polydenylated region. A potential polyadenylation signal (AATAAA) is found at positions 4164-4169. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human SULU1.

Multiple EST fragments match the 5′end (N27153) as well as the 3′ end (R90908) of the human SULU1 gene.

cDNA Cloning and Characterization of Murine SULU3

The murine SULU3 cDNA sequence is represented by PCR fragment 2R92-1-6.

A murine SULU3 clone, G83-4-5, was first identified from a PCR screen with degenerate oligos, CCK4c and CCK4b, applied to sscDNA generated from murine day-12 embryos. Sequence analysis of the 473 bp insert identified a 471 ORF (157 amino acids) with the potential to encode a novel human STK related to the C. elegans SULU gene (GB: U32275) product. The antisense strand of G83-4-5 is identical at the nucleic acid level to the 5′UTR of the murine etsl protooncogenic transcription factor (GB:X53953). This homology is likely the result of a cloning artifact attached to the 5′-end of the database entry for murine etsl.

PCR clone 3R19-17-6 was isolated from human A549 cell sscDNA using primers 5′-CCCCCGGGAAACGATGACCA(SEQ ID NO:59) and 5′-AGCCGCTGCCCCTCCTCTACTGT(SEQ ID NO:60) derived from G75-30-30 and R99-43-11, respectively. The 3′ primer used in this PCR reaction misprimed leading to the formation of a truncated product. 3R19-17-6 contains a 1172 bp insert and an ORF of 1170 bp (390 amino acids).

PCR clone 2R92-1-6 was isolated from murine d8 embryo sscDNA using primers 5′-ACCGCAACATCGCCACCTACTAC(SEQ ID NO:61) and 5′-GATTGCTTTGTGCTCATTCTTTGG(SEQ ID NO:72) derived from the 5′ UTR of the etsl gene and the human EST AA234623, respectively. The latter (shown herein) encodes the C-terminus of human SULU3.2R92-1-6 contains a 2249 bp insert and an ORF of 2244 bp (748 amino acids).

The partial murine SULU3 cDNA (SEQ ID NO.21) is 2249 bp long and consists of a 2244 bp ORF (6-2249) flanked by a 5 bp (1-5) 5′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for murine SULU3.

One EST fragment (AA446022) matches the 3′ end of the partial murine SULU3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of Human SULU3

The human SULU3 cDNA sequence is composed of two overlapping clones, 2R90-22-1 and AA234623.

A human SULU3 clone, AA234623, was first identified using a Smith-Waterman search of the EST database with the C. elegans SULU gene (GB: U32275) as a query. Sequence analysis of the 2652 bp insert identified a 1185 bp ORF (395 amino acids) with the potential to encode the C-terminus of a novel human STK related to the C. elegans SULU gene product.

PCR clone 2R90-22-1 was isolated from human SKMel128 melanoma cell line sscDNA using primers 5′-TATTGAATTGGCGGAACGGAAG(SEQ ID NO:70) and 5′-TTGTTCTAAGAGTGCCCTCCG(SEQ ID NO:73) derived from the murine SULU3 2R92-1-6 clone and from AA234623, respectively. 2R92-1-6 contains a 1897 bp insert and an ORF of 1896 bp (632 amino acids).

The partial human SULU3 cDNA (SEQ ID NO.20) is 3824 bp long. Clone 2R90-22-1 spans positions 1-1897 and clone AA234623 spans positions 1173. The overlaping region between these clones exhibits 100% sequence identity. The human SULU3 cDNA contains a 2358 bp ORF (2-2359) flanked by a 1465 bp (2360-3824) 3′UTR followed by a 19 nucleotide polydenylated region. A potential polyadenylation signal (AATAAA) is found at positions 2602-2607. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine.

Multiple EST fragments (R02283) match the 3′end of the human SULU3 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of GEK2

The human GEK2 cDNA sequence is composed of three overlapping clones, AA459448, 3R25-48-1 and GEK2_h#3.

A human GEK2 clone, AA459448, was first identified using a Smith-Waterman search of the EST database with the human SLK gene (GB: AB002804) as a query. Sequence analysis of the 1286 bp insert identified a 1227 bp ORF (409 amino acids) with the potential to encode the N-terminus of a novel human STK related to the human SLK gene product. An additional Smith-Waterman search using the C-terminus of the SLK gene as a query yielded three additional EST's, AA323687, AA380492 and AA168869, that encode the C-terminal region of human GEK2.

PCR clone 2R98-41-17 was isolated from human testis sscDNA using primers 5′-AAGACCATGCCGTGCGCCG(SEQ ID NO:74) and 5′-ATTCCTTCAGGTTCTGGTTATGG(SEQ ID NO:75) derived from AA323687 and from AA380492, respectively. 2R98-41-17 contains a 851 bp insert and an ORF of 849 bp (283 amino acids).

PCR clone GEK2_h#3 was isolated from human sscDNA made from the 1123 tumor cell line using primers 5′-GCAGCAAGTGGAGAAGATGG(SEQ ID NO:109) and 5′-GGAAGCATCCCCAGAGCTGTAG(SEQ ID NO:110) derived from the sequence of clone 3R25-48-l and from the 3′ end of murine LOK (GB:D89728), respectively. GEK2_h#3 contains a 1042 bp insert and an ORF of 1041 bp (347 amino acids).

The full-length human GEK2 cDNA (SEQ ID NO:106) is 2962 bp long. Clone AA459448 spans positions 1-1286, clone 3R25-48-1 spans positions 1100-2449 and clone GEK2_h#3 spans positions 1920-2962. The overlapping regions between these clones exhibit 100% sequence identity.

The human GEK2 cDNA contains a 2904 bp ORF (59-2962) flanked by a 58 bp (1-58) 5′UTR. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human GEK2.

Multiple EST fragments (AA465671) match the 5′end of the sequence, but only one (AA380492) matches the 3′ end of the human GEK2 gene.

cDNA Cloning and Characterization of PAK4

The human PAK4 cDNA sequence is represented by clone SNB2#1.

A human PAK4 clone, R88460, was first identified using a Smith-Waterman search of the EST database with the human PAK gene (GB: U24152) as a query. Sequence analysis of the 2332 bp insert identified a 930 bp ORF (310 amino acids) with the potential to encode the C-terminus of a novel human STK related to the human PAK gene product.

cDNA clone SNB2#1 was isolated from human glioblastoma cell line SNB75 cDNA library using a probe derived from R88460. SNB2#1 contains a 3604 bp insert and an ORF of 2043 bp (681 amino acids).

The full-length human PAK4 cDNA (SEQ ID NO.27) is 3604 bp long and consists of a 2043 bp ORF (143-2185) flanked by a 142 bp (1-142) 5′UTR and a 1419 3′ UTR followed by a 22 nucleotide polydenylated region. A potential polyadenylation signal (AATTAAA) is found at positions 3582-3588. The sequence flanking the first ATG conforms to the Kozak consensus for an initiating methionine, and is believed to be the translational start site for human PAK4. The 3′ UTR of the PAK4 gene contains a GT dinucleotide repeat prone to undergo expansion based on the number of repeats found in clones SNB#1 and R88460, 32 and 23, respectively. Several neurologic disorders have been correlated with the expansion of di- or tri-nucleotide repeats similar to those found in the PAK4 sequence, suggesting PAK 4 may also be a disease target and that this repeat in its 3′UTR may serve as a diagnostic marker.

Multiple EST fragments (AA535791) match the 3′end of the human PAK4 gene, but at the time of filing, the inventors believe that none exist in GenBank or the EST database that match its 5′ end.

cDNA Cloning and Characterization of PAK5

The full-length human PAK5 cDNA sequence is composed of two overlapping clones, H450#1-1 and SNB8#5.

A human PAK5 clone, R18825, was first identified using a Smith-Waterman search of the EST database with the human PAK4 gene as a query. Sequence analysis of the 1248 bp insert identified a 420 bp ORF (140 amino acids) with the potential to encode the C-terminus of a novel human STK related to the human PAK4 gene product.

cDNA clone SNB8#5 was isolated from human SNB75 cDNA library using a probe derived from R18825. SNB2#1 contains a 2028 bp insert and an ORF of 1194 bp (398 amino acids).

The partial human PAK5 cDNA (SEQ ID NO.28) is 2028 bp long and consists of a 1194 bp ORF (2-1195) flanked by an 833 bp (1196-2028) 3′UTR followed by a 22 nucleotide polydenylated region. A potential polyadenylation signal (AATTAAA) is found at positions 2004-2010. Since the coding region is open throughout the 5′ extent of this sequence, this is apparently a partial cDNA clone lacking the N-terminal start methionine.

Clone H460#1-1 was isolated from a human lung H460 cDNA library using a probe derived from the partial SNB2#1 cDNA clone described above. Sequence analysis of the 2526 bp insert identified a 1773 bp ORF (592 amino acids) with the potential to encode a full-length PAK5.

The human PAK5 cDNA (SEQ ID NO:102) is 2,806 bp long and consists of a 1,773 bp ORF flanked by a 201 bp 5′ UTR and a 833 bp 3′ UTR. The sequence flanking the first ATG conforms to the Kozak consensus (Kozak, M., Nucleic Acids Res. 15, 8125-8148 (1987)) for an initiating methionine, and is believed to be the translational start site for PAKS.

PAK5 shares 99% sequence identity over 2795 bp to a recent database entry, AF005046. These sequences are presumed to be from the same gene, with minor polymorphic variations.

Example 2 Expression Analysis of Mammalian STE20-Related Protein Kinases

Materials and Methods

Northern Blot Analysis

Northern blots were prepared by running 10 μg total RNA isolated from 60 human tumor cell lines (HOP-92, EKVX, NCI-H23, NCI-H226, NCI-H322M, NCI-H460, NCI-H522, A549, HOP-62, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, IGROVi, SKOV-3, SNB-19, SNB-75, U251, SF-268, SF-295, SF-539, CCRF-CEM, K-562, MOLT-4, HL-60, RPMI 8226, SR, DU-145, PC-3, HT-29, HCC-2998, HCT-116, 5W620, Cob 205, HTC15, KM-12, UO-31, SN12C, A498, CaKil, RXF-393, ACHN, 786-0, TK-10, LOX IMVI, Malme-3M, SK-MEL-2, SK-MEL-5, SK-MEL-28, UACC-62, UACC-257, M14, MCF-7, MCF-7/ADR RES, Hs578T, MDA-MB-231, MDA-MB-435, MDA-N, BT-549, T47D), from 22 human adult tissues (thymus, lung, duodenum, colon, testis, brain, cerebellum, cortex, salivary gland, liver, pancreas, kidney, spleen, stomach, uterus, prostate, skeletal muscle, placenta, mammary gland, bladder, lymph node, adipose tissue), and 2 human fetal normal tissues (fetal liver, fetal brain), on a denaturing formaldehyde 1.2% agarose gel and transferring to nylon membranes.

Filters were hybridized with random primed [α³²P]dCTP-labeled probes synthesized from the inserts of several of the STE20-related kinase genes. Hybridization was performed at 42° C. overnight in 6× SSC, 0.1% SDS, 1× Denhardt's solution, 100 μg/mL denatured herring sperm DNA with 1-2×10⁶ cpm/mL of ³²P-labeled DNA probes. The filters were washed in 0.1× SSC/0.1% SDS, 65° C., and exposed on a Molecular Dynamics phosphorimager.

Quantitative PCR Analysis

RNA was isolated from a variety of normal human tissues and cell lines. Single stranded cDNA was synthesized from 10 μg of each RNA as described above using the Superscript Preamplification System (GibcoBRL). These single strand templates were then used in a 25 cycle PCR reaction with primers specific to each clone. Reaction products were electrophoresed on 2% agarose gels, stained with ethidium bromide and photographed on a UV light box. The relative intensity of the STK-specific bands were estimated for each sample.

DNA Array Based Expression Analysis

Plasmid DNA array blots were prepared by loading 0.5 μg denatured plasmid for each STE20-related kinase on a nylon membrane. The [α³²P]dCTP labeled single stranded DNA probes were synthesized from the total RNA isolated from several human immune tissue sources or tumor cells (thymus, dendrocytes, mast cells, monocytes, B cells (primary, Jurkat, RPMI8226, SR), T cells (CD8/CD4+, TH1, TH2, CEM, MOLT4), K562 (megakaryocytes). Hybridization was performed at 42° C. for 16 hours in 6× SSC, 0.1% SDS, 1× Denhardt's solution, 100 μg/mL denatured herring sperm DNA with 10⁶ cpm/mL of [α³²P]dCTP labeled single stranded probe. The filters were washed in 0.1× SSC/0.1% SDS, 65° C., and exposed for quantitative analysis on a Molecular Dynamics phosphorimager.

Results

Distribution of STE20-Related Gene Transcripts in Normal Tissues and Tumor Cell Lines

ZC1, ZC2, and ZC3 RNA expression was analyzed by quantitative PCR from multiple human normal tissues, cultured primary epithelial and endothelial cells, and tumor cell lines. The results are summarized in Tables 1 and 2, with relative expression values ranging from 0 (undetectable) to 23 (very strong). An “x” refers to sample not tested. ZC1, ZC2, and ZC3 were all expressed at very low levels in most normal human tissues, however ZC1 and ZC2 were more abundant in cultured epithelial cells and ZC3 in normal kidney and breast tissue.

Expression of these 3 genes was also examined in a panel of human tumor cell lines representing a diverse sampling of tumor types (Table 2). ZC1 and ZC2 showed strong expression in cell lines from most melanomas and renal tumors and from some non-small cell lung cancers and colon tumors. ZC3 expression was consistently lower in the tumor cell lines except for high expression in most breast cancers and leukemias. The robust overexpression ZC1, ZC2, and ZC3 in tumor cells versus normal tissues may provide an attractive target for oncology drug development.

Expression of all the novel STE20-related kinases was examined in a panel of human immune tissues/cells by hybridization to a DNA array blot containing plasmids encoding each of these genes. STLK2 was broadly expressed in all 14 immune samples, whereas STLK4 and PAK4 were highly expressed in a subset of 6-7 of the samples (Table 3). Several other kinases (SULU3, ZC4, KHS2) had more restricted expression, while others were expressed in only a single immune source (STLK3, thymus; ZC1, dendrocytes; ZC3, monocytes; PAK5, mast cells and MOLT4), and several more were absent from all the immune sources assayed (GEK2, SULU1, ZC2, STLK5). These expression patterns were quite distinct among members of the same subfamily (i.e., ZC1, ZC2, ZC3 and ZC4, or PAK1, PAK2, PAK3, PAK4, PAK5). This analysis suggests that some of these kinases may be candidate targets for various immune disorders, and that some, which are more broadly expressed, may mediate functions vital to the basic biology of most proliferating cells.

TABLE 1 ZC1, ZC2 and ZC3 Expression in Normal Human Tissues and Cells Sample NORMAL ZC1 ZC2 ZC3 Brain Tiss 2.8 0.6 0.9 Duod Tiss 3.8 1.5 0.3 Heart Tiss 1.2 0.3 0.0 Kidney Tiss 0.7 0.0 7.0 Lung Tiss 1.6 0.2 0.0 Pancreas Tiss 2.0 0.4 2.5 Placenta Tiss 1.4 0.0 0.0 Sal gl. Tiss 3.0 0.3 3.2 Sk mus. Tiss 2.3 0.1 0.1 Spleen Tiss 0.4 0.0 x Stomach Tiss 0.8 0.0 0.0 Thymus Tiss 3.5 0.4 1.5 Cereb Tiss 2.8 1.1 4.4 Liver Tiss 1.8 0.0 0.4 Uterus Tiss 1.6 0.0 1.4 Prostate Tiss 1.4 0.0 1.6 Testis Tiss x x 5.8 f Brain Tiss x x 3.1 Mam gl Tiss x x 7.2 HCAEC ENDO 1.0 0.0 0.0 HMVEC-d ENDO 0.7 0.0 0.4 HMVEC-L ENDO 2.2 1.6 1.8 HPAEC ENDO 9.3 5.3 6.4 HMEC EPI 4.1 2.3 1.9 RPTEC EPI 3.6 2.2 0.2 HRCE EPI 5.3 3.5 1.3 HSAE EPI 0.9 3.3 4.8

TABLE 2 ZC1, ZC2 and ZC3 Expression in Tumor Cell lLnes Sample Origin ZC1 ZC2 ZC3 Sample Origin ZC1 ZC2 ZC3 HOP-92 Lung 9.3 7.2 3.3 HCC-2998 Colon 2.4 3.8 3.0 EKVX Lung 10.7 3.7 3.5 HCT 116 Colon 2.2 2.1 5.4 NCI-H23 Lung 5.8 6.3 4.1 SW-620 Colon 7.8 12.1 3.1 NCI-H226 Lung 6.5 6.8 3.3 COLO 205 Colon 9.1 16.2 3.0 NCI-H322M Lung 3.5 5.8 4.9 HCT-15 Colon 13.8 4.9 2.5 NCI-H460 Lung 4.5 3.7 2.9 KM-12 Colon 7.0 13.2 3.1 NCI-H522 Lung 4.7 3.3 4.6 UO-31 Colon 10.4 10.6 0.9 A549/ATCC Lung 3.8 3.6 4.1 SN12C Renal 8.1 3.4 2.8 HOP-62 Lung 4.3 3.8 4.2 A498 Renal 6.2 3.1 2.9 OVCAR-3 Ovary 2.9 3.1 1.5 Caki-1 Renal 9.2 14.4 2.3 OVCAR-4 Ovary 3.3 1.0 3.8 RXF 393 Renal 10.6 4.8 2.8 OVCAR-5 Ovary 2.6 3.6 2.2 ACHN Renal 9.3 6.0 3.9 OVCAR-8 Ovary 3.6 2.0 4.7 786-0 Renal 8.8 15.6 5.6 IGROV1 Ovary 3.8 1.7 3.2 TK-10 Renal 20.9 21.2 5.0 SK-OV-3 Ovary 4.9 0.0 3.5 LOX IMVI Mel 2.3 2.4 3.3 SNB-19 CNS 5.1 5.4 4.2 Malme-3M Mel x x 2.2 SNB-75 CNS 2.5 0.9 0.7 SK-MEL-2 Mel 15.7 14.1 2.9 U251 CNS 1.5 1.2 0.6 SK-MEL-5 Mel 7.9 7.0 0.0 SF-268 CNS 5.8 2.7 3.0 SK-MEL-28 Mel 16.5 23.1 0.0 SF-295 CNS 6.4 1.1 3.2 UACC-62 Mel 12.1 18.3 5.3 SF-539 CNS 5.1 2.9 4.3 UACC-257 Mel 10.8 9.4 6.2 CCRF-CEM Leuk 3.4 2.7 3.1 M14 Mel 4.4 0.9 7.9 K-562 Leuk 4.1 6.3 4.3 MCF7 Breast 4.8 1.3 7.7 MOLT-4 Leuk 7.1 3.4 4.2 MCF-7/ADR Breast 8.8 3.4 7.7 HL-60 Leuk x x 0.4 Hs 578T Breast 6.9 2.6 5.7 RPMI 8226 Leuk 0.5 0.2 1.4 MDA-MB-231 Breast 5.7 1.9 6.4 SR Leuk 3.5 7.2 5.4 MDA-MB-435 Breast 4.8 6.7 9.1 DU-145 Pro x x 3.4 MDA-N Breast 7.3 6.3 9.1 PC-3 Pro x x 3.4 BT-549 Breast 3.6 1.9 8.0 HT-29 Colon 2.4 5.9 6.6 T-47D Breast 0.4 12.3 9.3

TABLE 3 STE20-related kinase expression in a human immune panel Dendro- Mast Mono- B CD8+ CEM MOLT4 JURKAT RPMI8226 SR K562 KINASE thymus cytes cells cytes cells CD4+ TH1 TH2 (T cell) (T cell) (B cell) (B cell) (B cell) (MO) GEK2 350 350 350 350 350 350 350 350 350 350 350 350 350 350 SULU1 350 350 350 350 350 350 350 350 350 350 350 350 350 350 SULU3 350 350 350 350 12149 350 5115 350 350 350 350 350 350 350 STLK2 117770 13771 27620 92036 18305 39109 5408 3564 47236 53262 47605 22560 65936 30390 STLK3 8624 350 350 350 350 350 350 350 350 350 350 350 350 350 STLK4 8524 350 350 350 350 8685 5642 350 3648 350 26772 1570 350 350 STLK5 xxx xxx xxx xxx 350 350 350 xxx 350 350 350 xxx 350 350 ZC1 350 3377 350 350 350 350 350 350 350 350 350 350 350 350 ZC2 350 350 350 350 350 350 350 350 350 350 350 350 350 350 ZC3 350 350 350 20156 350 350 350 350 350 350 350 350 350 350 ZC4 xxx xxx xxx xxx 350 350 350 xxx 1094 7813 14945 xxx 350 6385 KHS2 8766 2508 350 56575 350 350 350 350 350 350 350 350 350 350 PAK4 32658 7684 3729 100948 350 350 350 1604 350 10246 350 3229 350 350 PAK5 350 350 4905 350 350 350 350 350 350 12672 350 350 350 350

Transcript size from Northern data Kinase (kb) STLK2 3.8 STLK4 5.0 ZC1 6.9/4.7 ZC2 6.0/8.0 ZC4 5 KHS2 4.4 SULU1 4.5 SULU3 10.0 GEK2 5.5 PAK4 4.8 PAK5 3.5

STLK2 is widely expressed; the highest expression levels were found in placenta, spleen and PBL.

STLK4 is also widely expressed in normal tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis, ovary, small intestine, colon, and peripheral blood lymphocytes. STLK4 was also detected in Jurkat T cells.

ZC1 is highly overexpressed in the following human cancer cell lines: HOP-92, EKVX, NCI-H23, NCI-H226, NCI-H322M, NCI-H522, A549, HOP-62 (lung); OVCAR-3, OVCAR-4, OVCAR-5 (ovary); SNB-19, U251, SF-268, SF-295, SF-539 (CNS); K-562, RPMI-8226 (leukemia); DU-145, PC-3 (prostate); HT-29, HCC-2998, HCT-116, SW620, COLO-205, HCT-15, KM-12 (colon); UO-31, CAKi-1, RXF-393, 786-0, TK-10 (renal); LOXIMVI, Malme-3M, SK-MEL-2, SK-MEL-28, UACC-62, UACC-257, M14 (melanoma); and MCF-7, MCF-7/ADR, HIS 578T, MDA-MB-231, MDA-MB-431, MDA-N, BT-549, T-47D (breast).

ZC2 is expressed in brain and testis. It is highly overexpressed in the following human cancer cell lines: TK-10 (renal); SK-MEL-28, UACC-62 (melanoma); T47D (breast).

Moderate expression in HOP92 (lung); OVCAR4, IGROV1 (ovary); DNB75, U251 (brain); K-562 (leukemia); and COL0205 (colon).

SULU1 is overexpressed in the following human cancer cell lines: HOP-92, EKVX, NCI-H23, NCI-H226, NCI-H322M, NCI-H522, A549, HOP-62 (lung); OVCAR-3, OVCAR-4, OVCAR-5, SK-OV-3 (ovary); SNB-19, U251, SF-268, SF-295, SF-539 (CNS); K-562, RPMI-8226 (leukemia); DU-145, PC-3 (prostate); HT-29, HCC-2998, HCT-116, SW620,. COLO-205, HCT-15, KM-12 (colon); UO-31, CAKi-1, RXF-393, 786-0, TK-10 (renal); LOX, IMVI, Malme-3M, SK-MEL-2, SK-MEL-28, UACC-62, UACC-257, M14 (melanoma); MCF-7, MCF-7/ADR, HIS 578T, MDA-MB-231, MDA-MB-431, MDA-N, BT-549, T-47D (breast)

SULU3 showed a broad pattern of expression in the normal tissue panel of RNAs.

GEK2 was expressed in spleen, thymus and testis. Expression was high in the cell lines RBL-2H3 and H441.

PAK4 was expressed in the normal tissues: brain, testis and prostate, and in the human cancer cell lines: HNCI-H23 (lung); OVCAR-3 (ovary); SNB-19, U251 (CNS); RPMI-8226 (leukemia); DU-145 (prostate); COLO-205, HCT-15 (colon).

PAK5 showed weak expression levels in the normal tissues: brain, testes, bladder, colon, adrenal medulla, spleen, fetal liver, breast, cerebral cortex, cerebellum, thymus, salivary gland, lung, stomach, duodenum, uterus, prostate, skeletal muscle and placenta. PAK5 was overexpressed in the human cancer cell lines: HOP-92, EKVX, NCI-H23, NCI-H226, NCI-H322M, NCI-H522, A549, HOP-62 (lung); OVCAR-3, OVCAR-4, OVCAR-5, SK-OV-3 (ovary); SNB-19, U251, SF-268, SF-295, SF-539 (CNS); K-562, RPMI-8226 (leukemia); DU-145, PC-3 (prostate); HT-29, HCC-2998, HCT-116, SW620,. COLO-205, HCT-15, KM-12 (colon); UO-31, CAKi-1, RXF-393, 786-0, TK-10 (renal); LOXIMVI, Malme-3M, SK-MEL-2, SK-MEL-28, UACC-62, UACC-257, M14 (melanoma); MCF-7, MCF-7/ADR, HIS 578T, MDA-MB-231, MDA-MB-431, MDA-N, BT-549, T-47D (breast).

Example 3 STE20-Related Protein Kinase Gene Expression Vector Construction

Materials and Methods

Expression Vector Construction

Several expression constructs were generated for some of the human STE20-related cDNAs including: a) full-length clones in a PcDNA expression vector; b) a GST-fusion construct containing the catalytic domain of the novel STE20-related kinase fused to the C-terminal end of a GST expression cassette; and c) a full-length clone containing a Lys to Ala (K to A) mutation at the predicted ATP binding site within the kinase domain, inserted in the pcDNA vector.

The “K to A” mutants of the STE20-related kinase might function as dominant negative constructs, and will be used to elucidate the function of these novel STKs.

Results

Constructs for ZC1, ZC2, ZC3, SULU1, SULU3, PAK4 and PAK5 have been generated.

Numerous additional constructs have been generated for the various STE20-subfamily kinases, including full length, kinase inactive and tagged versions. In addition, the following three constructs were designed for specific applications based on their unique domain structure:

Construct 1: SULU1-coiled-coil2

Vector: pGEX-4T

Insert: Coiled-coil2

Sequence: Amino acids 752-898

Purpose: phage display

Result: Interacts with GEK2 CC1

Construct 2: SULU3-coiled-coil2

Vector: pGEX4T

Insert: coiled-coil 2 domain fused to GST

Sequence range of insert: amino acids 802-898 of SEQ

Purpose: phage display

Result: Interacts with coiled—coiled region of human SLK

Construct 3: PAK5 Dominant Negative

Vector: pCAN5

Insert: Full length coding sequence of human PAK5

containing the following mutation: K350,351A (Lys at aa positions 350 and 351 changed to Ala).

Purpose: to determine role of human PAK5 kinase activity in cell growth and transformation.

Result: Interferes with Ras transformation.

Example 4 Generation of Specific Immunoreagents to STE20-Related Protein Kinases

Materials and Methods

Specific immunoreagents were raised in rabbits against KLH- or MAP-conjugated synthetic peptides corresponding to the human STE20-related kinases. C-terminal peptides were conjugated to KLH with glutaraldehyde, leaving a free C-terminus. Internal peptides were MAP-conjugated with a blocked N-terminus. Additional immunoreagents can also be generated by immunizing rabbits with the bacterially expressed GST-fusion proteins containing the cytoplasmic domains of each novel STK.

The various immune sera are first tested for reactivity and selectivity to recombinant protein, prior to testing for endogenous sources.

Western Blots

Proteins in SDS PAGE are transferred to immobilon membrane. The washing buffer is PBST (standard phosphate-buffered saline pH 7.4+0.1% triton×100). Blocking and antibody incubation buffer is PBST +5% milk. Antibody dilutions varied from 1:1000 to 1:2000.

Results

Three SULU1 antisera (against both 539A (SEQ ID NO: 79) and 540A (SEQ ID NO: 78)) and two SULU3 antisera (542A) (SEQ ID NO: 81) reacted specifically with the peptide antigens. Antisera binding was competable with peptide. Experiments with extracts from cells transfected with epitope-tagged SULU1 and SULU3 genes are underway.

Antisera against the PAK4 C-terminal peptide 554A (SEQ ID NO: 82) reacted with purified Gst-PAK4 and detected a protein of the correct molecular weight from tissue culture cells. Specific immunoprecipitation experiments are ongoing to determine the reactivity with native protein.

Similar immunization and antisera testing experiments are underway for each of the other novel STE20-kinases.

STE20-related protein kinase peptide immunogens and their specificity in recognizing endogenous protein by Western blots or immunoprecipitations. Aa Protein Sequence positions Conj West. IP STLK2 EKFQKCSADESP 405-416 KLH Y Y (SEQ ID NO:111) STLK4 SISNSELFPTTDPVGT 252-267 KLH Y Y (SEQ ID NO:112) SULU1 LDFPKEDYR 890-898 KLH Y Y (SEQ ID NO:113) SULU1 HGDPRPEPRPTQ 409-420 KLH Y Y (SEQ ID NO:114) SULU3 PSTNRAGSLKDPEC  2-14 KLH N ND (SEQ ID NO:115) SULU3 DPRTRASDPQSPPQVSRHK 411-429 KLH ND ND (SEQ ID NO:116) PAK4 CLVPLIQLYRKQTSTC 666-680 KLH ND Y (SEQ ID NO:117) PAK5 PLMRQNRTR 390-398 KLH Y Y (SEQ ID NO:118) PAK5 SGDRRRAGPEKRPKSS 148-163 KLH Y Y (SEQ ID NO:119) PAK5 (C) RRKSLVGTPYWMAPE 471-485 KLH Y ND (SEQ ID NO:120) ND = not done yet

STE20-related protein kinase GST fusion protein immunogens and their specificity in recognizing endogenous protein by Western blots or immunoprecipitations. Aa Protein domain positions West. IP ZC1 Coiled-coil/pro/B/C 350-867 Y Y ZC1 B 615-732 Y Y ZC2 Coiled-coil/pro/B 348-762 ND ND ZC2 B 658-762 Y Y PAK4 Nterm 252-426 ND ND PAK4 Kinase/Cterm 350-681 ND Y PAK5 A/Nterm  53-330 ND ND PAK5 A/Nterm  53-309 ND ND ND = not done yet

The 50 kD STLK2 protein was expressed highly in several hematopoietic cell lines including Jurkat, pGL10, Ramos, A20, WEHI-231, K562, HEL and freshly isolated thymocytes from C57/BL6 mice. High levels of STLK2 expression were also detected in several tumor cell lines including Calu6, Colo205, LS180, MDAM231 and A549.

The 160 kD ZC1 protein was detected in Jurkat T cells, Colo205, HCT116, RIE-1, 293T, MDAMB231, and SK-MEL28.

The 170 kD ZC2 protein was detected in SK-Mel28 and UACC-62.

Elevated levels of the 64 kD PAK5 protein were confirmed in the breast cancer cell lines MDA-231 and MCF-7, and in the lung cancer cell line A549.

Example 5 Recombinant Expression and Biological Assays for STE20-related Protein Kinases

Materials and Methods

Transient Expression of the Ste20-related Kinases in Mammalian Cells

The pcDNA expression plasmids (10 μg DNA/100 mm plate) containing the STE20-related kinase constructs are introduced into 293 cells with lipofectamine (Gibco BRL). After 72 hours, the cells are harvested in 0.5 mL solubilization buffer (20 mM HEPES, pH 7.35, 150 mM NaCl, 10% glycerol, 1% Triton X-100, 1.5 mM MgCl₂, 1 mM EGTA, 2 mM phenylmethylsulfonyl fluoride, 1 μg/mL aprotinin). Sample aliquots were resolved by SDS polyacrylamide gel electrophoresis (PAGE) on 6% acrylamide/0.5% bis-acrylamide gels and electrophoretically transferred to nitrocellulose. Non-specific binding was blocked by preincubating blots in Blotto (phosphate buffered saline containing 5% w/v non-fat dried milk and 0.2% v/v nonidet P-40 (Sigma)), and recombinant protein was detected using the various anti-peptide or anti-GST-fusion specific antisera.

In Vitro Kinase Assays

Three days after transfection with the STE20-related kinase expression contructs, a 10 cm plate of 293 cells was washed with PBS and solubilized on ice with 2 mL PBSTDS containing phosphatase inhibitors (10 mM NaHPO₄, pH 7.25, 150 mM NaCl, 1% Triton X-100, 0.5% deoxycholate, 0.1% SDS, 0.2% sodium azide, 1 mM NaF, 1 mM EGTA, 4 mM sodium orthovanadate, 1% aprotinin, 5 μg/mL leupeptin). Cell debris was removed by centrifugation (12000× g, 15 min, 4° C.) and the lysate was precleared by two successive incubations with 50 μL of a 1:1 slurry of protein A sepharose for 1 hour each. One-half mL of the cleared supernatant was reacted with 10 μL of protein A purified kinase-specific antisera (generated from the GST fusion protein or antipeptide antisera) plus 50 μL of a 1:1 slurry of protein A-sepharose for 2 hr at 4° C. The beads were then washed 2 times in PBSTDS, and 2 times in HNTG (20 mM HEPES, pH 7.5/150 mM NaCl, 0,1% Triton X-100, 10% glycerol).

The immunopurified kinases on sepharose beads were resuspended in 20 μL HNTG plus 30 mM MgCl₂, 10 mM MnCl₂, and 20 μCi [α³²P]ATP (3000 Ci/mmol). The kinase reactions were run for 30 min at room temperature, and stopped by addition of HNTG supplemented with 50 mM EDTA. The samples were washed 6 times in HNTG, boiled 5 min in SDS sample buffer and analyzed by 6% SDS-PAGE followed by autoradiography. Phosphoamino acid analysis was performed by standard 2D methods on ³²P-labeled bands excised from the SDS-PAGE gel.

Similar assays were performed on bacterially expressed GST-fusion constructs of the kinases.

ZC1 Assay buffer:

20 mM Tris pH 7.4, 200 mM NaCl, 0.5 mM DTT, 3 mM MgCl2, 0.3 mM MnCl2, 100 μM ³²PγATP.

Substrates: myelin basic protein (MBP) at 0.28 mg/mL and phosphorylated ZG1 peptide RTVGRRNTFIGT-PPYWMAPE(SEQ ID NO:121) at 17 μM (bold underlined residue shows site of phosphorylation).

At higher concentrations of MgCl₂ (3 mM), the activity of ZC1 (both full-length and recombinant kinase domain) is up to 10-fold greater towards exogenous substrate MBP. In contrast, the autophosphorylation and the phosphorylation of the activation loop peptide substrate are both inhibited. Mn++ does not inhibit the autophosphorylation and the peptide phosphorylation by the truncated kinase domain form. However, both the MBP phosphorylation, Mn++− preferring activity AND the autophosphorylating, Mg++− preferring activity are eliminated with mutation of the ATP-binding lysine in ZC1 (Lys54Ala) indicating that both activities are attributable to the ZC1 kinase domain.

SULU1 Assay Buffer:

This buffer is identical to that for ZC1, except for 5 mM MgCl2. Under these conditions, other STE20 family members (PAK4, ZC1) were inhibited for autophosphorylation and required reducing the [Mn] to <0.3 mM for an efficient autophosphorylation reaction.

Substrates: MBP, phosvitin, or α-casein at 0.28 mg/mL.

PAK4, PAK5 Assay Buffer:

20 mM Hepes pH 7.2, 130 mM KCl, 10 mM MgCl2, 1 mM NaF, 20 mM B-glycerolphosphate, 0.5 mM DTT, 50 μM ATP, 0.5 μCi ³²PyATP.

Substrates: MBP at 0.28 mg/mL and peptide substrates derived from PAK5 activation loop at 2.5 μM.

STLK2 Assay Buffer:

Similar to that described above, except for the inclusion of 5 mM MgCl₂, 5 mM MnCl₂ and 5 μCi ³²PγATP.

Transformation (PAK Experiments)

Low-passage NIH3T3 fibroblasts displaying normal morphology (flat, non-refractile cellular morphology), as well as low rates of spontaneous transformation, were used in transformation assays. NIH3T3 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% (v/v) fetal calf serum, penicillin (100 U/mL) and streptomycin (100 U/mL) and kept in an humidified incubator at 37° C. and 5% CO₂.

Cells were transfected with DNA-lipid complexes. As per manufacturer instructions, lipofectamine was utilized to transfect NIH3T3 cells. All transfections were with equal amounts of plasmid DNA (DNA from the appropriate expression vector without insert was used to give equivalent amounts of DNA per transfection). 1 μg of activated allele of H-Ras was co-transfected with increasing amounts of various alleles of PAK5.

Foci were scored after 3 weeks by fixing 10 min in 10% methanol, 10% acetic acid for 10 min, followed by staining with 0.4% (w/v) crystal violet in 10% methanol for 10 min, and washing with deionized water and drying at room temperature.

Transfections, Stimulations, and Luciferase Assays (ZC1 Experiments)

Cells (10⁷) were transiently transfected by electroporation using a Gene Pulser (Bio-Rad Labs) with the setting of 960_F and 250 V. 20-40 hours later, transfected cells (about 10⁵) were stimulated with various stimuli. After a 6-hour stimulation, cells were lysed, and luciferase activities were measured using the MicroLumatPlus (EG&G Berthold). (J. Exp. Med. 183:611-620, 1996, hereby incorporated by reference herein in its entirety including any drawings, tables, or figures.)

Results

Protein expression and kinase activity of novel STE20- related protein kinases Observed In vitro Endogenous size Predicted Kinase Kinase Protein (kD) Size (kD) activity activity STLK2  50  46 y y STLK4  55  50 y ND ZC1 160 140 y y ZC2 170 150 y y KHS2 ND 101 ND ND SULU1 119 105 y y SULU3 140 115 ND y PAK4  80  75 y y PAK5  64  64 y y

ZC1: Regulation of Kinase Activity

ZC1 is constitutively active as a full-length kinase when expressed either in vitro (TNT rabbit reticulocyte system) or in NIH 3T3, 293T, or H1299 tissue culture cells. The endogenously expressed kinase is also active when immunoprecipitated from carcinoma cell lines.

ZC1 Signaling Pathways

Using human leukemic T cell line Jurkat as a model system, the impact of cotransfected wild-type ZC1 on the activation of two reporter genes, RE/AP-luciferase and NFκB luciferase, was examined. RE/AP is a composite in the IL-2 gene promoter containing both a NFκB-like site and an AP-1 site.

Optimal activation of both RE/AP-luciferase and NFκB-luciferase reporter genes in Jurkat T cells requires signals generated from stimulation of both T cell receptor and the costimulator receptor CD28. Cotransfection of wild-type ZC1 with either the RE/AP-luciferase or the NFκB-luciferase reporter results in the activation of RE/AP or NFκB when costimulated with the anti-T cell receptor monoclonal antibody or the pharmacological reagents PMA and ionomycin that bypass proximal T cell receptor. No activation was seen when costimulated with an anti-CD28 monoclonal antibody.

These results suggest that wild-type ZC1, when overexpressed, was replacing a CD28-specific signal to activate RE/AP or NFκB. These results imply that ZC1 is involved in the CD28 signaling pathway. Since NFκB is one of the major pathways also activated by the pro-inflammatory cytokine TNF-α signaling, it is also likely that ZC1 may be a component in the TNF-α signaling pathways.

PAK5: Design of Specific Peptide Substrates

To aid in the development of in vitro kinase assays for screening small molecule libraries to identify specific inhibitors, the search for specific peptide substrates for PAK5 was undertaken.

The rationale used to design such peptides is based on the hypothesis that upon binding activated small G protein, PAK5 undergoes a conformational change that results in derepression of its kinase activity followed by autophosphorylation on the activation loop resulting in a fully active kinase. The site of autophosphorylation for related family members has been identified by biochemical and/or genetic means (e.g. Wu, C, et al. J.Biol.Chem 270:15984-15992 and Szczepanowska, et al. Proc.Natl.Acad.Sci 94, 8503-8508, 1997). Specific peptide substrates for PAKS were designed from the sequence of the activation loop of this kinase.

An activation loop PAK5 peptide phosphorylated on the Thr residue of the TPY motif served as a high-affinity substrate for PAK5.

PAK5 activation loop peptides as kinase substrates Peptide SEQ # Kinase Sequence Aa ID Kinase substrate Notes 1 PAK5 (C) RRKSLVGTPYWMAPE 471-485 120 PAK5 yes Equally well as MBB 2 PAK5 (C) RRKSLVG T PYWMAPE 471-485 120 PAK5 yes High Km for PAK5 (1-10 μM) 3 PAK5 (C) RRK S LVGTPYWMAPE 471-485 120 PAK5 no S is the site of phosphorylation 4 KHS1 KRKSFIGTPYWMAPE 171-185 122 PAK5 yes Similiar to peptide 1 5 STLK2 KRNTFVGTPFWMAPE 175-189 123 PAK5 poor 6 SULU1 PANSFVGTPYWMAPE 174-188 124 PAK5 poor 7 ZC1 RRNTFIGTPYWMAPE 184-198 125 PAK5 poor 8 ZC1 RRNTFIG T PYWMAPE 184-198 126 PAK5 poor Better than 7 9 STLK4 RNKVRKTFVGTPCWMAPE 66-83 127 PAK5 poor 10  PAK5 (C) RRKSLVGTPYWMAPE 471-485 120 PAK4 yes Same Km as phosph. by PAK5 Note: underlined/bold reside was phosphorylated

PAK5: Transformation

Transformation of low-passage NIH3T3 cells by ras in the presence or absence of various alleles of PAK5 showed that the dominant negative, kinase-dead allele of PAK5 was able to block ras transformation of NIH3T3 cells. Thus, PAK5 activity is required for ras transformation of NIH3T3 cells. Inhibition of PAK5 activity may have therapeutic value as an anti-proliferative agent for treating cancer.

PAK4 and PAK5: Interaction with Cdc42

PAK 4 interacts with CDC42 small G-protein but not Rac, RhoA, or Ras as determined by co-transfection of recombinant genes and detection by kinase assays. PAK5 also interacts with Cdc42. Coding sequences of activated alleles of small G proteins (ras, Cdc42, Rac, Rho) tagged with a Myc epitope were transiently expressed in 293T cells, various alleles of ³⁵S-labeled PAK5 tagged with HA epitope were expressed in vitro with the reticulocyte (TNT) system.

Example 6 Chromosomal Localization of Ste2O-Related Protein Kinases

Materials and Methods

STE20 protein kinases STLK3, STLK4, ZC1, ZC2, ZC3, KHS2, SULU1, PAK4, and PAK5 were mapped using the GeneBridge 4 Radiation Hybrid Panel, RHO2.05 (Research Genetics). The GeneBridge 4 Panel consists of 91 hybrid panel samples, in addition to one human positive control (HFL), and one hamster negative control (A23). The standard reaction conditions used to test and conduct PCR reactions using the GeneBridge 4 Panel are available from Research Genetics.

Oligonucleotide sequences (all 5′ to 3′) used for PCR mapping were:

STLK3: CTCCCATTTCCTAGCAAAATCA, (SEQ ID NO:128) AGAGGCAGTATTGTCAGATGTA (SEQ ID NO:129) STLK4: CCACACATGCGTATCTCTGTTG, (SEQ ID NO:130) TTGCTAGAATTCACATCAGGTACA (SEQ ID NO:131) ZC1: ATCCCTGGATCACACTGCTTCT, (SEQ ID NO:132) CAAGGTGTTCTTTGCCTCTGTT (SEQ ID NO:133) ZC2: AGATGGACTGTACTGGGAGGG, (SEQ ID NO:134) AGAAGAGCACTTGGCACTTATC (SEQ ID NO:135) ZC3: CATCATGAACTGGTGACGGG, (SEQ ID NO:136) CCAGTGAAATCAAACCAGTAAAA (SEQ ID NO:137) SULU1: CAAAACCTGGCCGTCTCTTCTATT, (SEQ ID NO:138) ATTTGTGCTACTGGGATTCTGTG (SEQ ID NO:139) KHS2: GAATAGCGGTACCATGATAGAATA, (SEQ ID NO:140) TACCAAAAAGAGCCAAAAGTGTG (SEQ ID NO:141) PAK4: CTCAGTATTCTCTCCAAAGATTG, (SEQ ID NO:142) GATGTTCTCTCCATTCTGTAAAG (SEQ ID NO:143) PAK5: CATCACTGGAAGTCTGCAGTG, (SEQ ID NO:144) CAGGTGCAGTAGTCATTTGC (SEQ ID NO:145)

Positive reactions were assigned a score of “1”, negative reactions are assigned a score of “0”, and ambiguous reactions are assigned a score of “2”. Results were submitted to the Whitehead Institute (www@genome.wi.mit.edu) for position analysis. Chromosomal localizations for ZC4, SULU3, STLK2, STLK5 and STLK6 were available publicly (for example, from Unigene). The chromosomal locations of GEK2 and STLK7 have not been determined.

STLK2_h Xq25-27.1 (Public) STLK3 2q31.3 (Sugen) STLK4_h 3p22.3-p22.2 (Sugen) STLK5_h 17q23.2-24.2 (Public) STLK6_h 2g32.2-q33.3 (Public) STLK7_h NA ZC1_h 2p11.2 (Sugen) ZC2_h 3q26.31-3q26.32 (Sugen) ZC3_h 17p13.2-13.3 (Sugen) ZC4_h Xq22 (Public) KHS2_h 2p22-2p22.2 (Sugen) SULU1_h 12q24.21 (Sugen) SULU3_h 17p11.2 (Public) GEK2_h NA PAK4_h 15q14 (Sugen) PAK5_h 19q13.2-q13.3 (Sugen)

Many of the STE 20 kinases were mapped to regions associated with various human cancers, as shown below.

The regions were also cross-checked with the Mendalian Inheritance in Man database, which tracks genetic information for many human diseases, including cancer. References for association of the mapped sites with chromosomal abnormalities found in human cancer can be found in: Knuutila, et al., Am J Pathol, 1998, 152:1107-1123, hereby incorporated herein be reference in its entirety including any figures, tables, or drawings.

Association of these mapped regions with other diseases is documented in the Online Mendalian Inheritance in Man (OMIM).

STLK2 h, Xq25-27.1, (Public)

Osteosarcoma, Xq25-qter, 2 of 31.

Lymphoproliferative syndrome, X-linked (OMIM No. 308240) human STLK3, 2q31.3, (Sugen)

Squamous cell carcinoma of Head and Neck, 3 of 30.

STLK4 h, 3p22.3-p22.2, (Sugen)

Mantle cell lymphoma 3pl4-p22 1 of 27

Squamous cell carcinoma of Head and Neck 3p22-p24 1 of 14

Cardiomyopathy, dilated (OMIM 601154)

STLK5 h, 17q23.2-24.2, (Public)

Cervical cancer, 17q, 1 of 30

Gastroesophageal junction adenocarcinoma xenograft, 17q, 1 of 5

Breast carcinoma, 17ql2-qter, 1 of 16

Bladder carcinoma, 17q22-q23, 1 of 14

Breast carcinoma, 17q22-q25, 8 of 101

Non-small cell lung cancer, 17q24-q25, 6 of 50

Testis, 17q24-qter, 2 of 11

Malignant peripheral nerve sheath tumors, 17q24-qter, 5 of 7

Alzheimer disease, susceptibility to (OMIM 106180)

STLK6 h, 2q32.2-q33.3, (Public)

Non-small cell lung cancer, 2q31-q32, 1 of 50

Squamous cell carcinoma of Head and Neck, 2q31-q33, 3 of 30

Small cell lung cancer, 2q32-q35, 1 of 22

ZC1 h, 2p11.2, (Sugen)

non-small cell lung cancer, 2pter-ql3, 1 of 10

non-small cell lung cancer, 2pter-q21, 1 of 10

Pulmonary alveolar proteinosis, congenital (OMIM 178640).

ZC2 h, 3q26.31-3q26.32, (Sugen)

Non-small cell lung cancer, 3q26.1-q26.3, 26 of 103

Cervical cancer, 3q26.1-q27, 4 of 30

Small cell lung cancer, 3q26.3-qter, 3 of 35

Squamous cell carcinoma of Head and Neck, 3q26.3-qter, 3 of 13

Marginal zone B-cell lymphoma, 3q26-q27, 1 of 25

Parosteal osteosarcoma, 3q26-q28, 1 of 1

Gastrointestinal stromal tumor, 3q26-q29, 1 of 16

Mantle cell lymphoma, 3q26-q29, 1 of 5

ZC3 h 17pl3.2-13.3 (Sugen)

Malignant fibrous histiocytoma of soft tissue, 17p, 2 of 58

Leiomyosarcoma, 17p, 7 of 29

Non-small cell lung cancer, 17p, 1 of 50

ZC4 h, Xq22, (Public)

Diffuse large cell lymphoma, Xq22-ter, 1 of 32

Deafness, X-linked 1, progressive. (OMIM 304700).

KHS2 h, 2p22-2p22.2, (Sugen)

Synovial sarcoma, 2p21-q14, 1_of_(—)67

Follicular lymphoma, 2p22-p24, 1_of_(—)46

Colorectal cancer, hereditary, nonpolyposis, type 1,

Ovarian cancer (MSH2, COCA1, FCC1). (OMIM 120435).

SULU1 h, 12q24.21 (Sugen)

Neuroglial tumors, 12q22-qter, 1_of_(—)15

Gastroesophageal junction adenocarcinoma, 12q23-qter, 1 of 5.

Non-small cell lung cancer, 12q24.1-24.3, 2 of 50.

SULU3 h 17p11.2 (Public)

Malignant fibrous histiocytoma of soft tissue, 17p, 2_of_(—)58

Leiomyosarcoma, 17p, 7_of_(—)29

non-small cell lung cancer, 17p, 1_of_(—)50

Diffuse large cell lymphoma, 17p11.2, 1_of_(—)32

Osteosarcoma, 17p11.2-pl2, 4_of_(—)31

PAK4 h: 15ql4 (Sugen)

Schizophrenia, (OMIM 118511).

PAK5 h: 19ql3.2-ql3.3 (Sugen)

Follicular lymphoma, 19ql3, 1 of 46*

Mantle cell lymphoma, 19ql3, 1 of 5

Hepatocellular carcinoma, 19ql3.1, 2 of 50

Small cell lung cancer, 19ql3.1, 10 of 35

Breast carcinoma, 19ql3.1-qter, 1 of 33

cervical cancer, 19ql3.1-qter, 1 of 30

Testis, 19ql3.1-qter, 1 of 11

Chondrosarcoma, 19ql3.2, 1 of 29

Malignant fibrous histiocytoma of soft tissue, 19q13.2-qter, 2 of 58

Non-small cell lung cancer, 19qcen-q13.3, 6 of 104

Example 7 Demonstration of Gene Amplification by Southern Blotting

Materials and Methods

Nylon membranes were purchased from Boehringer Mannheim. Denaturing solution contains 0.4 M NaOH and 0.6 M NaCl. Neutralization solution contains 0.5 M Tris-HCL, pH 7.5 and 1.5 M NaCl. Hybridization solution contains 50% formamide, 6× SSPE, 2.5× Denhardt's solution, 0.2 mg/mL denatured salmon DNA, 0.1 mg/mL yeast tRNA, and 0.2% sodium dodecyl sulfate. Restriction enzymes were purchased from Boehringer Mannheim. Radiolabeled probes were prepared using the Prime-it II kit by Stratagene. The beta actin DNA fragment used for a probe template was purchased from Clontech.

Genomic DNA was isolated from 20 different tumor cell lines: MCF-7, MDA-MB-231, Calu-6, A549, HCT-15, HT-29, Colo 205, LS-180, DLD-1, HCT-116, PC3, CAPAN-2, MIA-PaCa-2, PANC-1, AsPc-1, BxPC-3, OVCAR-3, SKOV3, SW 626 and PA-1, and from two normal cell lines: human mammary epithelial cells and human umbilical vein endothelial cells.

A 10 μg aliquot of each genomic DNA sample was digested with EcoR I restriction enzyme and a separate 10 μg sample was digested with Hind III restriction enzyme. The restriction-digested DNA samples were loaded onto a 0.7% agarose gel and, following electrophoretic separation, the DNA was capillary-transferred to a nylon membrane by standard methods (Sambrook, J. et al (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory).

PAK5 Amplicon:

A 600 base pair fragment (EcoR 1- Sac I) of the PAK5 gene was used as a template for a radiolabeled DNA probe which was hybridized to the blots at 42° C. for 48 hours in hybridization solution using standard methods (supra). The blots were exposed to a phosphorimager screen for 4 days, then scanned and analyzed using a Molecular Dynamics Storm 840 phosphorimager. The relative mass and gene copy number values of the PAKS DNA fragments were calculated from the band density values obtained. The blots were re-hybridized with a radiolabeled probe copied from a fragment of human beta actin DNA and developed as above to confirm the sample mass loading equivalency.

Results

The PAK5 gene was determined to exhibit 3-fold amplification compared to the normal DNA copy number in PANC-1 (pancreatic epithelioid carcinoma) and OVCAR-3 (ovarian adenocarcinoma) human cell lines, and approximately 2 times the normal copy number in the BxPC-3 (primary pancreatic adenocarcinoma) human cell line.

Similar Southern analyses can be performed for other STE20 kinases.

Example 8 Detection of Protein—Protein Interaction through Phage Display

Materials and Methods

Phage display provides a method for isolating molecular interactions based on affinity for a desired bait. cDNA fragments cloned as fusions to phage coat proteins are displayed on the surface of the phage. Phage(s) interacting with a bait are enriched by affinity purification and the insert DNA from individual clones is analyzed.

T7 Phage Display Libraries

All libraries were constructed in the T7Selectl-lb vector (Novagen) according to the manufacturer's directions.

Bait Presentation

Protein domains to be used as baits were generated as C-terminal fusions to GST and expressed in E. coli. Peptides were chemically synthesized and biotinylated at the N-terminus using a long chain spacer biotin reagent.

Selection

Aliquots of refreshed libraries (10¹⁰-10¹² pfu) supplemented with PanMix and a cocktail of E. coli inhibitors (Sigma P-8465) were incubated for 1-2 hrs at room temperature with the immobilized baits. Unbound phage was extensively washed (at least 4 times) with wash buffer.

After 3-4 rounds of selection, bound phage was eluted in 100 μL of 1% SDS and plated on agarose plates to obtain single plaques.

Identification of Insert DNAs

Individual plaques were picked into 25 μL of 10 mM EDTA and the phage was disrupted by heating at 70° C. for 10 min. 2 μL of the disrupted phage were added to 50 μL PCR reaction mix. The insert DNA was amplified by 35 rounds of thermal cycling (94° C., 50 sec; 50° C., 1 min; 72° C., 1 min).

Composition of Buffer

10× PanMix

5% Triton X100

10% non-fat dry milk (Carnation)

10 mM EGTA

250 mM NaF

250 μg/mL Heparin (sigma)

250 μg/mL sheared, boiled salmon sperm DNA (sigma)

0.05% Na azide

Prepared in PBS

Wash Buffer

PBS supplemented with:

0.5% NP-40

25 μl g/mL heparin

PCR Reaction Mix

PCR reaction mix 1.0 mL 10x PCR buffer (Perkin-Elmer, with 15 mM Mg) 0.2 mL each dNTPs (10 mM stock) 0.1 mL T7UP primer (15 pmol/μL) (SEQ ID NO:146) GGAGCTGTCGTATTCCAGTC 0.1 mL T7DN primer (15 pmol/μL) AACCCCTCAAGACCCGTTTAG (SEQ ID NO:147) 0.2 mL 25 mM MgCl₂ or MgSO₄ to compensate for EDTA Q.S. to 10 mL with distilled water Add 1 unit of Taq polymerase per 50 μL reaction

LIBRARY: T7 Selectl-H441

Results

Phage display baits and interactors Sequence CDNA Range Bait Domain Aa SEQ ID library Interactor & SEQ ID SULU1 Coiled- 752-898 22 H441 GEK2 cc 677-820 coil2 dom (1) SEQ #26 SULU3 Coiled- 755-898 23 H441 SLK M83780 coil2 isoform (1) SULU1 cc1 also interacted to a lesser extent with the coiled-coil domain of an SLK isoform.

The phage display data suggest potential interactions of SULU3 with SLK and SULU1 with GEK2 through their coiled-coil domains. Therefore two members of the SULU subfamily of STE20 kinases interact with two members of a separate STE20 family, the prototype being SLK.

These results suggest a specificity in the interaction, and imply that these STE20 kinases may interact with each other through homo- and hetero-dimerization. Alternatively SULU-related kinases could act immediately up- or down-stream of the SLK-related kineses in a signaling cascade.

One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The molecular complexes and the methods, procedures, treatments, molecules, specific compounds described herein are presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention.

All patents and publications mentioned in the specification are indicative of the levels of those skilled in the art to which the invention pertains.

The invention illustratively described herein suitably may be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of” and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed.

In particular, although some formulations described herein have been identified by the excipients added to the formulations, the invention is meant to also cover the final formulation formed by the combination of these excipients. Specifically, the invention includes formulations in which one to all of the added excipients undergo a reaction during formulation and are no longer present in the final formulation, or are present in modified forms.

In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. For example, if X is described as selected from the group consisting of bromine, chlorine, and iodine, claims for X being bromine and claims for X being bromine and chlorine are fully described.

Other embodiments are within the following claims.

                   #             SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 155 <210> SEQ ID NO 1 <211> LENGTH: 3268 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 1 taacagccca cctcctagcc ccgggctacg cgccgccagc ccagtaaccc ca #cttttgtg     60 tgtcctccca ggccccgatc gaaaagcctg ggagggccgc cgaactaccc cc #ggagggag    120 gagccagtcc gaacccaagg cgccaccgcc gcagaagcgg agcgaggcag ca #ttcgcctc    180 catggcccac tcgccggtgg ctgtccaagt gcctgggatg cagaataaca ta #gctgatcc    240 agaagaactg ttcacaaaat tagagcgcat tgggaaaggc tcatttgggg aa #gttttcaa    300 aggaattgat aaccgtaccc agcaagtcgt tgctattaaa atcatagacc tt #gaggaagc    360 cgaagatgaa atagaagaca ttcagcaaga aataactgtc ttgagtcaat gt #gacagctc    420 atatgtaaca aaatactatg ggtcatattt aaaggggtct aaattatgga ta #ataatgga    480 atacctgggc ggtggttcag cactggatct tcttcgagct ggtccatttg at #gagttcca    540 gattgctacc atgctaaagg aaattttaaa aggtctggac tatctgcatt ca #gaaaagaa    600 aattcaccga gacataaaag ctgccaatgt cttgctctca gaacaaggag at #gttaaact    660 tgctgatttt ggagttgctg gtcagctgac agatacacag attaaaagaa at #acctttgt    720 gggaactcca ttttggatgg ctcctgaagt tattcaacag tcagcttatg ac #tcaaaagc    780 tgacatttgg tcattgggaa ttactgctat tgaactagcc aagggagagc ca #cctaactc    840 cgatatgcat ccaatgagag ttctgtttct tattcccaaa aacaatcctc ca #actcttgt    900 tggagacttt actaagtctt ttaaggagtt tattgatgct tgcctgaaca aa #gatccatc    960 atttcgtcct acagcaaaag aacttctgaa acacaaattc attgtaaaaa at #tcaaagaa   1020 gacttcttat ctgactgaac tgatagatcg ttttaagaga tggaaggcag aa #ggacacag   1080 tgatgatgaa tctgattccg agggctctga ttcggaatct accagcaggg aa #aacaatac   1140 tcatcctgaa tggagcttta ccaccgtacg aaagaagcct gatccaaaga aa #gtacagaa   1200 tggggcagag caagatcttg tgcaaaccct gagttgtttg tctatgataa tc #acacctgc   1260 atttgctgaa cttaaacagc aggacgagaa taacgctagc aggaatcagg cg #attgaaga   1320 actcgagaaa agtattgctg tggctgaagc cgcctgtccc ggcatcacag at #aaaatggt   1380 gaagaaacta attgaaaaat ttcaaaagtg ttcagcagac gaatccccct aa #gaaactta   1440 ttattggctt ctgtttcata tggacccaga gagccccacc aaacctacgt ca #agattaac   1500 aatgcttaac ccatgagctc catgtgcctt ttggatcttt gcaacactga ag #atttggaa   1560 gaagctatta aactattttg tgatggcgtt tatcatttta tattttgaaa gg #attatttt   1620 gtaaggaata acttttaata ctatagtttc acctgtattc tagtaaatgt tg #agacaccg   1680 ttttgctttt aagtatccct atttcttaag ttacgaggat gaataccttt ca #cattttga   1740 tctttagttg actctacagt catgaaacat acaggtcttt caaagtcatt ct #caatattc   1800 agcttttgta aattatcaag cttcaaaaag ctttttttta aaaaaaaaaa ca #tgcatatt   1860 ctaaaaatga ctattggtgg ggaggtgtaa ataagtcata ccttcttaaa ac #agaaaatt   1920 taagtaaagt cttttaaatg aaacctgtaa aagtattgac tcttctacca ag #ttggtatg   1980 atattccagg cagctcaatg attatcacat ttgagaccct gtgtttgaag ca #tttacagg   2040 caatgtacag caacagaggt acctcttggt gtatagtatt tacattctct tt #taggtaga   2100 agaggcaatt ttacccttat ttcacatggt tagaaattta aagcaagatc at #ttacccaa   2160 ggataggtgt ttggtaatgt tgaaggagtt agtctggctt catgttttac at #cttcaact   2220 aaaatcccat actatctgct tggatttgga gagccaaaaa ataaagctga tt #gtcatgtg   2280 attaaatatc tgatcaacag gtatgaatat aacttaaatc agcatatttt tg #ccatggta   2340 ataaattgtc ctataaacta tttatatatt tttgttcttc ataattatca ct #aataagca   2400 tcagtttgtt gtttttaaaa ggatatttaa gtgagcattt tctagttcat at #gaaaataa   2460 ccatagtaca ggatgatttc tgtccacaca aaggttaaat tagattgcac ag #ttaatttt   2520 cacttatatt tatggtacta ttatgtgggt gatgcctttt tcttttaagc cc #agtacata   2580 tattatgcct gcctaagttc tgaactgggg ctgtatttca gtagttgtag aa #ttattgat   2640 atttagtttt gatagctaat gtttaattgt ttggatctgc acagtttggt tt #ttgcacaa   2700 aagtcattta aaaaaatctg agtaattgtc aaatattaaa agaaagatat tc #ttcctgta   2760 aggaatacag tttttagtca aagtggccat tacatcctct ttttaattta ca #taatacag   2820 atacttgaga aagttgttgt ggtgttgtat gccaagaaaa ttctttttat tg #gtgcctat   2880 attgtaacaa ttatttttaa tgcattgtat tttgaagtaa cggttcagtt aa #atttttca   2940 cctgctgtgt aactgaaaca caattacagt ttataatcat ctgtagaagt ct #ggagataa   3000 ttttgcaact catgttatgg gttaaatgaa tatttttgta aaagtaaaag ca #acaaattt   3060 ataaattgat tatttgaaac tttacaacac aattgcatcc caaatacaaa tt #gtattgct   3120 tattcattat agctattcgt cctgtaatct gtttctaggt gaagcatact cc #agtgtttt   3180 aggggttttg aaaataaata tttaaatttc acagtcaaaa aaaaaaaaaa aa #aaaaaaaa   3240 aaaaaaaaaa aaaaaaaaaa aaaaaaaa          #                   #           3268 <210> SEQ ID NO 2 <211> LENGTH: 3030 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 2 gacagcagcg ccggccccgg cagctcccgc ggccccggcc ccggccccgg cc #ccggcccc     60 ggcggcacag gctgtcggct ggcccatctg cagggacgcg tacgagctgc ag #gaggttat    120 cggcagtgga gctactgctg tggttcaggc agccctatgc aaacccaggc aa #gaacgtgt    180 agcaataaaa cggatcaact tggaaaaatg ccagaccagt atggatgaac ta #ttaaaaga    240 aattcaagcc atgagtcagt gcagccatcc caacgtagtg acctattaca cc #tcttttgt    300 ggtcaaagat gaactttggc tggtcatgaa attactaagt ggaggttcaa tg #ttggatat    360 cataaaatac attgtcaacc gaggagaaca caagaatgga gttctggaag ag #gcaataat    420 agcaacaatt cttaaagagg ttttggaagg cttagactat ctacacagaa ac #ggtcagat    480 tcacagggat ttgaaagctg gtaatattct tctgggtgag gatggttcag ta #caaatagc    540 agattttggg gtaagtgcgt tcctagcaac agggggtgat gttacccgaa at #aaagtaag    600 aaaaacattc gttggcaccc catgttggat ggctcctgaa gtcatggaac ag #gtgagagg    660 ctatgacttc aaggctgaca tgtggagttt tggaataact gccattgaat ta #gcaacagg    720 agcagcgcct tatcacaaat atcctcccat gaaagtgtta atgttgactt tg #caaaatga    780 tccacccact ttggaaacag gggtagagga taaagaaatg atgaaaaagt ac #ggcaagtc    840 ctttagaaaa ttactttcac tgtgtcttca gaaagatcct tccaaaaggc cc #acagcagc    900 agaactttta aaatgcaaat tcttccagaa agccaagaac agagagtacc tg #attgagaa    960 gctgcttaca agaacaccag acatagccca aagagccaaa aaggtaagaa ga #gttcctgg   1020 gtcaagtggt caccttcata aaaccgaaga cggggactgg gagtggagtg ac #gacgagat   1080 ggatgagaag agcgaagaag ggaaagcagc tttttctcag gaaaagtcac ga #agagtaaa   1140 agaagaaaat ccagagattg cagtgagtgc cagcaccatc cccgaacaaa ta #cagtccct   1200 ctctgtgcac gactctcagg gcccacccaa tgctaatgaa gactacagag aa #gcttcttc   1260 ttgtgccgtg aacctcgttt tgagattaag aaactccaga aaggaactta at #gacatacg   1320 atttgagttt actccaggaa gagatacagc agatggtgta tctcaggagc tc #ttctctgc   1380 tggcttggtg gatggtcacg atgtagttat agtggctgct aatttacaga ag #attgtaga   1440 tgatcccaaa gctttaaaaa cattgacatt taagttggct tctggctgtg at #gggtcgga   1500 gattcctgat gaagtgaagc tgattgggtt tgctcagttg agtgtcagct ga #tgtatgtc   1560 ccttgatgtc accctgatct gtcatgcccc accgccaccc ctactccctt ca #accctccc   1620 tctttctgcc catttcctcc caccccctca ctcccatttc ctagcaaaat ca #gaagattg   1680 tgaagaggcc ggcttcaaca aaatgggata aaaaaataat tttttaaaac tt #acaacact   1740 ccgagttctg ctttattctc tagcaatcca cagtacaaga acaagcaaat gc #cacagctg   1800 cacgactgtt gctcattttt ccaaaagcta tttaatattc ttagcaatca at #ttggatat   1860 cccttaagtg aaaagaatct gaaatacact caggtggtct tatttattgg ca #acaaaagg   1920 aattttctat ccagaagcct atttctcctt tcattgttgt tatttctgtt at #aatacttt   1980 aattgtacat ctgacaatac tgcctctttt atgttgtatt tagaaattaa ta #tacttata   2040 aaattaagat ttattagcca aacttgaatt ctagttttaa aactgactgt ga #attttatt   2100 tttcatatat ttatgcatta cacaccttag ctataagaaa aaaagggttt tg #attatatg   2160 cttcttgcag ttaatctcgt tatttaaaca aaaagttttg ggtctatctt tg #gagtattt   2220 gtaacttcta aattttgaaa tgactgaatt aggaatttgg atgcttattc tt #ttagtctg   2280 tttgcctaaa aaccaattta caatctgact gtctcttggg agagggaggt gc #cttgcaaa   2340 ctttcacatt aagaatgtgc ctgaggctgc tttactctgg aatagtctca ga #tctaaaat   2400 ttcctctata taaggtggca tatgttaagt tttgcttcat tggaccgttt ag #aatgctat   2460 gtaaaatgtt gccattctgt tagattgcta actatatacc catctctgat tt #ggctctcc   2520 ttaagtgata ggatttgtta ttctaaaggt gataaacttg aaaatatcag aa #tctgagtt   2580 ttacttgaaa ttttgcagaa tacccaggtg gagtgaaaat tggaagggtt tt #gtgcaatg   2640 actaaaaggt aaaacgctgt taaggttcaa gaatcaatac tttcaaccca ag #tagccctc   2700 tgcttgactg tatattatgg aactagtaaa ccttaggatt ttgaaaattg ga #gtctaatc   2760 tttcaaggag gtgggctccc aggatggtac cattgctctt tcctagctaa cc #ctagatat   2820 ggcagctctt taatgtactt caaaaagcaa atatatatta ctaaggaaaa aa #agttattt   2880 ataattgcct tgtcataatt gttaaggtgt tctagagcca tttgcataca at #ttaatgta   2940 atttcattcc attctattgt ttacacaacg attactcgaa gatgactgca aa #ggtaaaag   3000 gaaaataaaa gtgtattgca caatgaaaaa          #                   #         3030 <210> SEQ ID NO 3 <211> LENGTH: 3857 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (22) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (81) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (122) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 3 caaaagtgga gtcctagatg antctaccat tgctacgata ctccgagaag ta #ctggaagg     60 gctggaatat ctgcataaaa ntggacagat ccacagagat gtgaaagctg ga #aacattct    120 tnttggagaa gatggctcag tacagatttc agactttggg gttagtgctt tt #ttagcaac    180 tggtggtgat attacccgaa ataaagtgag aaagaccttt gttggcaccc ct #tgttggat    240 ggcacctgaa gttatggaac aggtccgtgg ttatgatttc aaagctgata tt #tggagttt    300 tggaattaca gcaattgaat tggctacagg ggcggctcct tatcataaat at #ccaccaat    360 gaaggtttta atgctgacac tgcagaacga tcctccttct ttggaaactg gt #gttcaaga    420 taaagaaatg ctgaaaaaat atggaaaatc atttagaaaa atgatttcat tg #tgccttca    480 aaaagatcca gaaaaaagac caacagcagc agaactatta aggcacaaat tt #ttccagaa    540 agcaaagaat aaagaatttc ttcaagaaaa aacattgcag agagcaccaa cc #atttctga    600 aagagcaaaa aaggttcgga gagtaccagg ttccagtggg cgtcttcata ag #acagagga    660 tggaggctgg gagtggagtg atgatgaatt tgatgaagaa agtgaggaag gg #aaagcagc    720 aatttcacaa ctcaggtctc cccgagtgaa agaatcaata tcaaattctg ag #ctctttcc    780 aacaactgat cctgtgggta ctttgctcca agttccagaa cagatctctg ct #catctacc    840 tcagccagct gggcagattg ctacacagcc aactcaagtc tctctcccac cc #accgcaga    900 gccagcaaaa acagctcagg ctttgtcttc aggatcaggt tcacaagaaa cc #aagatccc    960 aatcagtcta gtactaagat taaggaattc caaaaaagaa ctaaatgata tt #cgatttga   1020 atttactcct gggagagata cagcagaggg tgtctctcag gaactcattt ct #gctggcct   1080 ggtcgacgga agggatttag taatagtggc agctaatttg cagaaaattg tg #gaagaacc   1140 tcagtcaaat cgatctgtca ctttcaaact ggcatctggt gtcgaaggct ca #gatattcc   1200 tgatgatggt aaactgatag gatttgccca gctcagcatc agctaaacca ca #accctgga   1260 agaggcggcc taaggagatt ccacacatgc gtatctctgt tgcttctatt gg #cctaaacc   1320 cactactgcc aaagaaccca gcaacaaacc tcccggctag gagctttaga ag #tctttatg   1380 ttcttcctgc catcattcct ccttttccca cagggaaaga aaagttggat ca #ctagtggc   1440 cagcatcccc agagttccgt tagtaaactt acttcatatg tcccctgtct tc #ctccatct   1500 gagaagtggc ccatgtgctt caaggcccag gagggagatc tgtcagctca tt #cttgcctt   1560 actccaatga tggcccaggt ggaaaagtag cagctgtatc gggcttcctc at #cctgcctg   1620 ttcccccaca cctgccagga tatggacatc ttgggatatc tctttaccac tg #aagtagaa   1680 ttgattgttc agctggagcc cagagaattt aatttaatgt tttttctttg ta #cctgatgt   1740 gaattctagc aacctttgtt aggaaaaagc acagcctcag atggaggcag cc #taaactgt   1800 gttcttgttt tgttcatggt gtttctaagc gttttgctga agctgctctc ag #gcaccccc   1860 ttcttcattg ctctctccag aaagggttgc tagccttaac ttcagctggt gc #aaaacatc   1920 tgactgtagc cgaacttcag ccatcagatc cttcaaagtg gaactttgga tt #gtttttac   1980 agacaacatc gagtaatggc ttgtaaatgt gaattttgcc agaggtggtt tt #tgaacagg   2040 aaaatcataa ttcatatcat tggagaagta tttattttca aatatcaaat tg #aagaaaaa   2100 ctcaatcctc ccatgaaaat cagttcgcct ggcctccaag tcgtgaggaa at #gggtatgc   2160 aaggctgaga tttctacagc aataaaggag acacacactg ggccagagag gc #ctgccttc   2220 tgcctgctct cctgcactga ccctttggag ggggtctctg tgtgctgaag ct #aactcaag   2280 atggaaagtg aaaccacatg tgccgtgacc tttaggtttt atgagtagac ag #tgttcatt   2340 tgattttcta cagaaataat ataaattatt ctttaggttt aaaaaagagc ac #tcataatg   2400 caatatgtga ataatcagtg aggttgattt ttcttttttc ctaccgtttc at #agtctttg   2460 tctaactgct agtaacccta ccgagtttta tatatgagtg ggatactcaa tc #tggcctta   2520 aaaagataca caaagatggg ctgtgggtcc ctggaaaggg ggagagttgc cc #tttacaga   2580 atcactcgag ccctttccag cactgttggt ctgatgaaca aggttgtttt ac #cttatttt   2640 ctcttggaac atatctgaaa accttcccca caaataactt gtcacacctt tt #gtttcatt   2700 ctgagtcttt agttttagtc atgggctttc ttcacctgct ctaggtgcaa ag #gcatgttg   2760 ggaaagagat ggatgttggg gaggaagaga ggagatggat ttcagttggg ag #ttaggagg   2820 agagtaggtg agatgatcag acaccggagt tcaacgtccc agcagtcttg gt #aaaaggag   2880 ggagcctgct gagccaggag ggagaaaaga agattgacca gcttgctaga aa #aatactta   2940 gcttttcttt ttcttttttt gtggaggggg gacggagagg aacaaggatg gg #gaggtagg   3000 aatgaggtat agaaaagaga tagcatcttc tttggcacaa gactagtggc tt #accgctta   3060 ccttagagtt ttgttttttt tttttcaaac ccatcaaaat ctacttattt at #gaatccaa   3120 ggggtggcag catcactctg ttctagcatt ctttgtggag atggtctggt gc #ctagctgg   3180 gagtgagcag cagcccatcc cctgttcact ttctctagcc catcattacc tg #tgaactgc   3240 agtggggcag tcatggcaaa tagaattggg ctggggtttc tccttctttt ca #gttcattg   3300 tttgccctgc taggaattag aagacagaca ccatgtccca ggacagtgtt ac #ttcttctg   3360 catgatgtgt ggtagactcc ctttgctggc ttgtgcagtg atactgagaa aa #tacatgaa   3420 cagaaactgc ccaggtggaa cagcacgtaa cctagtgagt gactgtactc ct #ttctagga   3480 atgctgattc agagtgcacc tctttgacta ggtcccagga tccccttgtc cc #tggagtag   3540 ggactaacta tagcacaaag taatatgtgc caatgctatt tgtgaaatgt tt #ggtctttc   3600 taaacgacta aaggatttgt tgggtttttg cttaagtttt gaaccaaatc ct #agagccag   3660 ctgataatat ttaataatct ggaggagaga ataatgatgt accaataagt gg #agattcct   3720 ccttatgatg tatgctaggt tatggaagat gtaaaatatt caactttttc ct #cctttttt   3780 tggactttgt attttactgc atgttttctt catttttaat caataaagag ta #aattgtca   3840 aaaaaaaaaa aaaaaaa              #                   #                   # 3857 <210> SEQ ID NO 4 <211> LENGTH: 1584 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 4 ctcatctgta cacacttcat ggatggcatg aatgagctgg cgattgctta ca #tcctgcag     60 ggggtgctga aggccctcga ctacatccac cacatgggat atgtacacag ga #gtgtcaaa    120 gccagccaca tcctgatctc tgtggatggg aaggtctacc tgtctggttt gc #gcagcaac    180 ctcagcatga taagccatgg gcagcggcag cgagtggtcc acgattttcc ca #agtacagt    240 gtcaaggttc tgccgtggct cagccccgag gtcctccagc agaatctcca gg #gttatgat    300 gccaagtctg acatctacag tgtgggaatc acagcctgtg aactggccaa cg #gccatgtc    360 ccctttaagg atatgcctgc cacccagatg ctgctagaga aactgaacgg ca #cagtgccc    420 tgcctgttgg ataccagcac catccccgct gaggagctga ccatgagccc tt #cgcgctca    480 gtggccaact ctggcctgag tgacagcctg accaccagca ccccccggcc ct #ccaacggt    540 gactcgccct cccaccccta ccaccgaacc ttctcccccc acttccacca ct #ttgtggag    600 cagtgccttc agcgcaaccc ggatgccagg cccagtgcca gcaccctcct ga #accactct    660 ttcttcaagc agatcaagcg acgtgcctca gaggctttgc ccgaattgct tc #gtcctgtc    720 acccccatca ccaattttga gggcagccag tctcaggacc acagtggaat ct #ttggcctg    780 gtaacaaacc tggaagagct ggaggtggac gattgggagt tctgagcctc tg #caaactgt    840 gcgcattctc cagccaggga tgcagaggcc acccagaggc ccttcctgag gg #ccggccac    900 attcccgccc tcctgggcag attgggtaga aaggacattc ttccaggaaa gt #tgactgct    960 gactgattgg gaaagaaaat cctggagaga tacttcactg ctccaaggct tt #tgagacac   1020 aagggaatct caacaaccag ggatcaggag ggtccaaagc cgacattccc ag #tcctgtga   1080 gctcaggtga cctcctccgc agaagagaga tgctgctctg gccctgggag ct #gaattcca   1140 agcccagggt ttggctcctt aaacccgagg accgccacct cttcccagtg ct #tgcgacca   1200 gcctcattct atttaacttt gctctcagat gcctcagatg ctataggtca gt #gaaagggc   1260 aagtagtaag ctgcctgcct cccttccctc agacctctcc ctcataattc ca #gagaaggg   1320 catttctgtc tttttaagca cagactaagg ctggaacagt ccatccttat cc #ctcttctg   1380 gcttgggccc tgacacctaa gtctttccca cggtttatgt gtgtgcctca tt #cctttccc   1440 accaagaatc catcttagcg cctcctgcca gctgccctgg tgctttctcc aa #gggccatc   1500 agtgtcttgc ctagcttgag ggcttaagtc cttatgctgt gttagtttcg tt #gtcagaac   1560 aaattaaaat tttcagagac gctg           #                   #              1584 <210> SEQ ID NO 5 <211> LENGTH: 416 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 5 Met Ala His Ser Pro Val Ala Val Gln Val Pr #o Gly Met Gln Asn Asn   1               5  #                 10  #                 15 Ile Ala Asp Pro Glu Glu Leu Phe Thr Lys Le #u Glu Arg Ile Gly Lys              20      #             25      #             30 Gly Ser Phe Gly Glu Val Phe Lys Gly Ile As #p Asn Arg Thr Gln Gln          35          #         40          #         45 Val Val Ala Ile Lys Ile Ile Asp Leu Glu Gl #u Ala Glu Asp Glu Ile      50              #     55              #     60 Glu Asp Ile Gln Gln Glu Ile Thr Val Leu Se #r Gln Cys Asp Ser Ser  65                  # 70                  # 75                  # 80 Tyr Val Thr Lys Tyr Tyr Gly Ser Tyr Leu Ly #s Gly Ser Lys Leu Trp                  85  #                 90  #                 95 Ile Ile Met Glu Tyr Leu Gly Gly Gly Ser Al #a Leu Asp Leu Leu Arg             100       #           105       #           110 Ala Gly Pro Phe Asp Glu Phe Gln Ile Ala Th #r Met Leu Lys Glu Ile         115           #       120           #       125 Leu Lys Gly Leu Asp Tyr Leu His Ser Glu Ly #s Lys Ile His Arg Asp     130               #   135               #   140 Ile Lys Ala Ala Asn Val Leu Leu Ser Glu Gl #n Gly Asp Val Lys Leu 145                 1 #50                 1 #55                 1 #60 Ala Asp Phe Gly Val Ala Gly Gln Leu Thr As #p Thr Gln Ile Lys Arg                 165   #               170   #               175 Asn Thr Phe Val Gly Thr Pro Phe Trp Met Al #a Pro Glu Val Ile Gln             180       #           185       #           190 Gln Ser Ala Tyr Asp Ser Lys Ala Asp Ile Tr #p Ser Leu Gly Ile Thr         195           #       200           #       205 Ala Ile Glu Leu Ala Lys Gly Glu Pro Pro As #n Ser Asp Met His Pro     210               #   215               #   220 Met Arg Val Leu Phe Leu Ile Pro Lys Asn As #n Pro Pro Thr Leu Val 225                 2 #30                 2 #35                 2 #40 Gly Asp Phe Thr Lys Ser Phe Lys Glu Phe Il #e Asp Ala Cys Leu Asn                 245   #               250   #               255 Lys Asp Pro Ser Phe Arg Pro Thr Ala Lys Gl #u Leu Leu Lys His Lys             260       #           265       #           270 Phe Ile Val Lys Asn Ser Lys Lys Thr Ser Ty #r Leu Thr Glu Leu Ile         275           #       280           #       285 Asp Arg Phe Lys Arg Trp Lys Ala Glu Gly Hi #s Ser Asp Asp Glu Ser     290               #   295               #   300 Asp Ser Glu Gly Ser Asp Ser Glu Ser Thr Se #r Arg Glu Asn Asn Thr 305                 3 #10                 3 #15                 3 #20 His Pro Glu Trp Ser Phe Thr Thr Val Arg Ly #s Lys Pro Asp Pro Lys                 325   #               330   #               335 Lys Val Gln Asn Gly Ala Glu Gln Asp Leu Va #l Gln Thr Leu Ser Cys             340       #           345       #           350 Leu Ser Met Ile Ile Thr Pro Ala Phe Ala Gl #u Leu Lys Gln Gln Asp         355           #       360           #       365 Glu Asn Asn Ala Ser Arg Asn Gln Ala Ile Gl #u Glu Leu Glu Lys Ser     370               #   375               #   380 Ile Ala Val Ala Glu Ala Ala Cys Pro Gly Il #e Thr Asp Lys Met Val 385                 3 #90                 3 #95                 4 #00 Lys Lys Leu Ile Glu Lys Phe Gln Lys Cys Se #r Ala Asp Glu Ser Pro                 405   #               410   #               415 <210> SEQ ID NO 6 <211> LENGTH: 516 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 6 Thr Ala Ala Pro Ala Pro Ala Ala Pro Ala Al #a Pro Ala Pro Ala Pro   1               5  #                 10  #                 15 Ala Pro Ala Pro Ala Ala Gln Ala Val Gly Tr #p Pro Ile Cys Arg Asp              20      #             25      #             30 Ala Tyr Glu Leu Gln Glu Val Ile Gly Ser Gl #y Ala Thr Ala Val Val          35          #         40          #         45 Gln Ala Ala Leu Cys Lys Pro Arg Gln Glu Ar #g Val Ala Ile Lys Arg      50              #     55              #     60 Ile Asn Leu Glu Lys Cys Gln Thr Ser Met As #p Glu Leu Leu Lys Glu  65                  # 70                  # 75                  # 80 Ile Gln Ala Met Ser Gln Cys Ser His Pro As #n Val Val Thr Tyr Tyr                  85  #                 90  #                 95 Thr Ser Phe Val Val Lys Asp Glu Leu Trp Le #u Val Met Lys Leu Leu             100       #           105       #           110 Ser Gly Gly Ser Met Leu Asp Ile Ile Lys Ty #r Ile Val Asn Arg Gly         115           #       120           #       125 Glu His Lys Asn Gly Val Leu Glu Glu Ala Il #e Ile Ala Thr Ile Leu     130               #   135               #   140 Lys Glu Val Leu Glu Gly Leu Asp Tyr Leu Hi #s Arg Asn Gly Gln Ile 145                 1 #50                 1 #55                 1 #60 His Arg Asp Leu Lys Ala Gly Asn Ile Leu Le #u Gly Glu Asp Gly Ser                 165   #               170   #               175 Val Gln Ile Ala Asp Phe Gly Val Ser Ala Ph #e Leu Ala Thr Gly Gly             180       #           185       #           190 Asp Val Thr Arg Asn Lys Val Arg Lys Thr Ph #e Val Gly Thr Pro Cys         195           #       200           #       205 Trp Met Ala Pro Glu Val Met Glu Gln Val Ar #g Gly Tyr Asp Phe Lys     210               #   215               #   220 Ala Asp Met Trp Ser Phe Gly Ile Thr Ala Il #e Glu Leu Ala Thr Gly 225                 2 #30                 2 #35                 2 #40 Ala Ala Pro Tyr His Lys Tyr Pro Pro Met Ly #s Val Leu Met Leu Thr                 245   #               250   #               255 Leu Gln Asn Asp Pro Pro Thr Leu Glu Thr Gl #y Val Glu Asp Lys Glu             260       #           265       #           270 Met Met Lys Lys Tyr Gly Lys Ser Phe Arg Ly #s Leu Leu Ser Leu Cys         275           #       280           #       285 Leu Gln Lys Asp Pro Ser Lys Arg Pro Thr Al #a Ala Glu Leu Leu Lys     290               #   295               #   300 Cys Lys Phe Phe Gln Lys Ala Lys Asn Arg Gl #u Tyr Leu Ile Glu Lys 305                 3 #10                 3 #15                 3 #20 Leu Leu Thr Arg Thr Pro Asp Ile Ala Gln Ar #g Ala Lys Lys Val Arg                 325   #               330   #               335 Arg Val Pro Gly Ser Ser Gly His Leu His Ly #s Thr Glu Asp Gly Asp             340       #           345       #           350 Trp Glu Trp Ser Asp Asp Glu Met Asp Glu Ly #s Ser Glu Glu Gly Lys         355           #       360           #       365 Ala Ala Phe Ser Gln Glu Lys Ser Arg Arg Va #l Lys Glu Glu Asn Pro     370               #   375               #   380 Glu Ile Ala Val Ser Ala Ser Thr Ile Pro Gl #u Gln Ile Gln Ser Leu 385                 3 #90                 3 #95                 4 #00 Ser Val His Asp Ser Gln Gly Pro Pro Asn Al #a Asn Glu Asp Tyr Arg                 405   #               410   #               415 Glu Ala Ser Ser Cys Ala Val Asn Leu Val Le #u Arg Leu Arg Asn Ser             420       #           425       #           430 Arg Lys Glu Leu Asn Asp Ile Arg Phe Glu Ph #e Thr Pro Gly Arg Asp         435           #       440           #       445 Thr Ala Asp Gly Val Ser Gln Glu Leu Phe Se #r Ala Gly Leu Val Asp     450               #   455               #   460 Gly His Asp Val Val Ile Val Ala Ala Asn Le #u Gln Lys Ile Val Asp 465                 4 #70                 4 #75                 4 #80 Asp Pro Lys Ala Leu Lys Thr Leu Thr Phe Ly #s Leu Ala Ser Gly Cys                 485   #               490   #               495 Asp Gly Ser Glu Ile Pro Asp Glu Val Lys Le #u Ile Gly Phe Ala Gln             500       #           505       #           510 Leu Ser Val Ser         515 <210> SEQ ID NO 7 <211> LENGTH: 414 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (7) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (27) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (41) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 7 Lys Ser Gly Val Leu Asp Xaa Ser Thr Ile Al #a Thr Ile Leu Arg Glu   1               5  #                 10  #                 15 Val Leu Glu Gly Leu Glu Tyr Leu His Lys Xa #a Gly Gln Ile His Arg              20      #             25      #             30 Asp Val Lys Ala Gly Asn Ile Leu Xaa Gly Gl #u Asp Gly Ser Val Gln          35          #         40          #         45 Ile Ala Asp Phe Gly Val Ser Ala Phe Leu Al #a Thr Gly Gly Asp Ile      50              #     55              #     60 Thr Arg Asn Lys Val Arg Lys Thr Phe Val Gl #y Thr Pro Cys Trp Met  65                  # 70                  # 75                  # 80 Ala Pro Glu Val Met Glu Gln Val Arg Gly Ty #r Asp Phe Lys Ala Asp                  85  #                 90  #                 95 Ile Trp Ser Phe Gly Ile Thr Ala Ile Glu Le #u Ala Thr Gly Ala Ala             100       #           105       #           110 Pro Tyr His Lys Tyr Pro Pro Met Lys Val Le #u Met Leu Thr Leu Gln         115           #       120           #       125 Asn Asp Pro Pro Ser Leu Glu Thr Gly Val Gl #n Asp Lys Glu Met Leu     130               #   135               #   140 Lys Lys Tyr Gly Lys Ser Phe Arg Lys Met Il #e Ser Leu Cys Leu Gln 145                 1 #50                 1 #55                 1 #60 Lys Asp Pro Glu Lys Arg Pro Thr Ala Ala Gl #u Leu Leu Arg His Lys                 165   #               170   #               175 Phe Phe Gln Lys Ala Lys Asn Lys Glu Phe Le #u Gln Glu Lys Thr Leu             180       #           185       #           190 Gln Arg Ala Pro Thr Ile Ser Glu Arg Ala Ly #s Lys Val Arg Arg Val         195           #       200           #       205 Pro Gly Ser Ser Gly Arg Leu His Lys Thr Gl #u Asp Gly Gly Trp Glu     210               #   215               #   220 Trp Ser Asp Asp Glu Phe Asp Glu Glu Ser Gl #u Glu Gly Lys Ala Ala 225                 2 #30                 2 #35                 2 #40 Ile Ser Gln Leu Arg Ser Pro Arg Val Lys Gl #u Ser Ile Ser Asn Ser                 245   #               250   #               255 Glu Leu Phe Pro Thr Thr Asp Pro Val Gly Th #r Leu Leu Gln Val Pro             260       #           265       #           270 Glu Gln Ile Ser Ala His Leu Pro Gln Pro Al #a Gly Gln Ile Ala Thr         275           #       280           #       285 Gln Pro Thr Gln Val Ser Leu Pro Pro Thr Al #a Glu Pro Ala Lys Thr     290               #   295               #   300 Ala Gln Ala Leu Ser Ser Gly Ser Gly Ser Gl #n Glu Thr Lys Ile Pro 305                 3 #10                 3 #15                 3 #20 Ile Ser Leu Val Leu Arg Leu Arg Asn Ser Ly #s Lys Glu Leu Asn Asp                 325   #               330   #               335 Ile Arg Phe Glu Phe Thr Pro Gly Arg Asp Th #r Ala Glu Gly Val Ser             340       #           345       #           350 Gln Glu Leu Ile Ser Ala Gly Leu Val Asp Gl #y Arg Asp Leu Val Ile         355           #       360           #       365 Val Ala Ala Asn Leu Gln Lys Ile Val Glu Gl #u Pro Gln Ser Asn Arg     370               #   375               #   380 Ser Val Thr Phe Lys Leu Ala Ser Gly Val Gl #u Gly Ser Asp Ile Pro 385                 3 #90                 3 #95                 4 #00 Asp Asp Gly Lys Leu Ile Gly Phe Ala Gln Le #u Ser Ile Ser                 405   #               410 <210> SEQ ID NO 8 <211> LENGTH: 274 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 8 Leu Ile Cys Thr His Phe Met Asp Gly Met As #n Glu Leu Ala Ile Ala   1               5  #                 10  #                 15 Tyr Ile Leu Gln Gly Val Leu Lys Ala Leu As #p Tyr Ile His His Met              20      #             25      #             30 Gly Tyr Val His Arg Ser Val Lys Ala Ser Hi #s Ile Leu Ile Ser Val          35          #         40          #         45 Asp Gly Lys Val Tyr Leu Ser Gly Leu Arg Se #r Asn Leu Ser Met Ile      50              #     55              #     60 Ser His Gly Gln Arg Gln Arg Val Val His As #p Phe Pro Lys Tyr Ser  65                  # 70                  # 75                  # 80 Val Lys Val Leu Pro Trp Leu Ser Pro Glu Va #l Leu Gln Gln Asn Leu                  85  #                 90  #                 95 Gln Gly Tyr Asp Ala Lys Ser Asp Ile Tyr Se #r Val Gly Ile Thr Ala             100       #           105       #           110 Cys Glu Leu Ala Asn Gly His Val Pro Phe Ly #s Asp Met Pro Ala Thr         115           #       120           #       125 Gln Met Leu Leu Glu Lys Leu Asn Gly Thr Va #l Pro Cys Leu Leu Asp     130               #   135               #   140 Thr Ser Thr Ile Pro Ala Glu Glu Leu Thr Me #t Ser Pro Ser Arg Ser 145                 1 #50                 1 #55                 1 #60 Val Ala Asn Ser Gly Leu Ser Asp Ser Leu Th #r Thr Ser Thr Pro Arg                 165   #               170   #               175 Pro Ser Asn Gly Asp Ser Pro Ser His Pro Ty #r His Arg Thr Phe Ser             180       #           185       #           190 Pro His Phe His His Phe Val Glu Gln Cys Le #u Gln Arg Asn Pro Asp         195           #       200           #       205 Ala Arg Pro Ser Ala Ser Thr Leu Leu Asn Hi #s Ser Phe Phe Lys Gln     210               #   215               #   220 Ile Lys Arg Arg Ala Ser Glu Ala Leu Pro Gl #u Leu Leu Arg Pro Val 225                 2 #30                 2 #35                 2 #40 Thr Pro Ile Thr Asn Phe Glu Gly Ser Gln Se #r Gln Asp His Ser Gly                 245   #               250   #               255 Ile Phe Gly Leu Val Thr Asn Leu Glu Glu Le #u Glu Val Asp Asp Trp             260       #           265       #           270 Glu Phe <210> SEQ ID NO 9 <211> LENGTH: 3798 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 9 gagaccatgg cgaacgactc tcccgcgaaa agtctggtgg acatcgacct ct #cctccctg     60 cgggatcctg ctgggatttt tgagctggtg gaagtggttg gaaatggcac ct #atggacaa    120 gtctataagg gtcgacatgt taaaacgggt cagttggcag ccatcaaagt ta #tggatgtc    180 actgaggatg aagaggaaga aatcaaactg gagataaata tgctaaagaa at #actctcat    240 cacagaaaca ttgcaacata ttatggtgct ttcatcaaaa agagccctcc ag #gacatgat    300 gaccaactct ggcttgttat ggagttctgt ggggctgggt ccattacaga cc #ttgtgaag    360 aacaccaaag ggaacacact caaagaagac tggatcgctt acatctccag ag #aaatcctg    420 aggggactgg cacatcttca cattcatcat gtgattcacc gggatatcaa gg #gccagaat    480 gtgttgctga ctgagaatgc agaggtgaaa cttgttgact ttggtgtgag tg #ctcagctg    540 gacaggactg tggggcggag aaatacgttc ataggcactc cctactggat gg #ctcctgag    600 gtcatcgcct gtgatgagaa cccagatgcc acctatgatt acagaagtga tc #tttggtct    660 tgtggcatta cagccattga gatggcagaa ggtgctcccc ctctctgtga ca #tgcatcca    720 atgagagcac tgtttctcat tcccagaaac cctcctcccc ggctgaagtc aa #aaaaatgg    780 tcgaagaagt tttttagttt tatagaaggg tgcctggtga agaattacat gc #agcggccc    840 tctacagagc agcttttgaa acatcctttt ataagggatc agccaaatga aa #ggcaagtt    900 agaatccagc ttaaggatca tatagatcgt accaggaaga agagaggcga ga #aagatgaa    960 actgagtatg agtacagtgg gagtgaggaa gaagaggagg aagtgcctga ac #aggaagga   1020 gagccaagtt ccattgtgaa cgtgcctggt gagtctactc ttcgccgaga tt #tcctgaga   1080 ctgcagcagg agaacaagga acgttccgag gctcttcgga gacaacagtt ac #tacaggag   1140 caacagctcc gggagcagga agaatataaa aggcaactgc tggcagagag ac #agaagcgg   1200 attgagcagc agaaagaaca gaggcgacgg ctagaagagc aacaaaggag ag #agcgggaa   1260 gctagaaggc agcaggaacg tgaacagcga aggagagaac aagaagaaaa ga #ggcgtcta   1320 gaggagttgg agagaaggcg caaagaagaa gaggagagga gacgggcaga ag #aagaaaag   1380 aggagagttg aaagagaaca ggagtatatc aggcgacagc tagaagagga gc #agcggcac   1440 ttggaagtcc ttcagcagca gctgctccag gagcaggcca tgttactgga gt #gccgatgg   1500 cgggagatgg aggagcaccg gcaggcagag aggctccaga ggcagttgca ac #aagaacaa   1560 gcatatctcc tgtctctaca gcatgaccat aggaggccgc acccgcagca ct #cgcagcag   1620 ccgccaccac cgcagcagga aaggagcaag ccaagcttcc atgctcccga gc #ccaaagcc   1680 cactacgagc ctgctgaccg agcgcgagag gtggaagata gatttaggaa aa #ctaaccac   1740 agctcccctg aagcccagtc taagcagaca ggcagagtat tggagccacc ag #tgccttcc   1800 cgatcagagt ctttttccaa tggcaactcc gagtctgtgc atcccgccct gc #agagacca   1860 gcggagccac aggttcctgt gagaacaaca tctcgctccc ctgttctgtc cc #gtcgagat   1920 tccccactgc agggcagtgg gcagcagaat agccaggcag gacagagaaa ct #ccaccagt   1980 attgagccca ggcttctgtg ggagagagtg gagaagctgg tgcccagacc tg #gcagtggc   2040 agctcctcag ggtccagcaa ctcaggatcc cagcccgggt ctcaccctgg gt #ctcagagt   2100 ggctccgggg aacgcttcag agtgagatca tcatccaagt ctgaaggctc tc #catctcag   2160 cgcctggaaa atgcagtgaa aaaacctgaa gataaaaagg aagttttcag ac #ccctcaag   2220 cctgctgatc tgaccgcact ggccaaagag cttcgagcag tggaagatgt ac #ggccacct   2280 cacaaagtaa cggactactc ctcatccagt gaggagtcgg ggacgacgga tg #aggaggac   2340 gacgatgtgg agcaggaagg ggctgacgag tccacctcag gaccagagga ca #ccagagca   2400 gcgtcatctc tgaatttgag caatggtgaa acggaatctg tgaaaaccat ga #ttgtccat   2460 gatgatgtag aaagtgagcc ggccatgacc ccatccaagg agggcactct aa #tcgtccgc   2520 cggactcagt ccgctagtag cacactccag aaacacaaat cttcctcctc ct #ttacacct   2580 tttatagacc ccagattact acagatttct ccatctagcg gaacaacagt ga #catctgtg   2640 gtgggatttt cctgtgatgg gatgagacca gaagccataa ggcaagatcc ta #cccggaaa   2700 ggctcagtgg tcaatgtgaa tcctaccaac actaggccac agagtgacac cc #cggagatt   2760 cgtaaataca agaagaggtt taactctgag attctgtgtg ctgccttatg gg #gagtgaat   2820 ttgctagtgg gtacagagag tggcctgatg ctgctggaca gaagtggcca ag #ggaaggtc   2880 tatcctctta tcaaccgaag acgatttcaa caaatggacg tacttgaggg ct #tgaatgtc   2940 ttggtgacaa tatctggcaa aaaggataag ttacgtgtct actatttgtc ct #ggttaaga   3000 aataaaatac ttcacaatga tccagaagtt gagaagaagc agggatggac aa #ccgtaggg   3060 gatttggaag gatgtgtaca ttataaagtt gtaaaatatg aaagaatcaa at #ttctggtg   3120 attgctttga agagttctgt ggaagtctat gcgtgggcac caaagccata tc #acaaattt   3180 atggccttta agtcatttgg agaattggta cataagccat tactggtgga tc #tcactgtt   3240 gaggaaggcc agaggttgaa agtgatctat ggatcctgtg ctggattcca tg #ctgttgat   3300 gtggattcag gatcagtcta tgacatttat ctaccaacac atatccagtg ta #gcatcaaa   3360 ccccatgcaa tcatcatcct ccccaataca gatggaatgg agcttctggt gt #gctatgaa   3420 gatgaggggg tttatgtaaa cacatatgga aggatcacca aggatgtagt tc #tacagtgg   3480 ggagagatgc ctacatcagt agcatatatt cgatccaatc agacaatggg ct #ggggagag   3540 aaggccatag agatccgatc tgtggaaact ggtcacttgg atggtgtgtt ca #tgcacaaa   3600 agggctcaaa gactaaaatt cttgtgtgaa cgcaatgaca aggtgttctt tg #cctctgtt   3660 cggtctggtg gcagcagtca ggtttatttc atgaccttag gcaggacttc tc #ttctgagc   3720 tggtagaagc agtgtgatcc agggattact ggcctccaga gtcttcaaga tc #ctgagaac   3780 ttggaattcc ttgtaact              #                   #                   #3798 <210> SEQ ID NO 10 <211> LENGTH: 4055 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 10 gctttcgggg aggtctatga gggtcgtcat gtcaaaacgg gccagcttgc ag #ccatcaag     60 gttatggatg tcacagggga tgaagaggaa gaaatcaaac aagaaattaa ca #tgttgaag    120 aaatattctc atcaccggaa tattgctaca tactatggtg cttttatcaa aa #agaaccca    180 ccaggcatgg atgaccaact ttggttggtg atggagtttt gtggtgctgg ct #ctgtcacc    240 gacctgatca agaacacaaa aggtaacacg ttgaaagagg agtggattgc at #acatctgc    300 agggaaatct tacgggggct gagtcacctg caccagcata aagtgattca tc #gagatatt    360 aaagggcaaa atgtcttgct gactgaaaat gcagaagtta aactagtgga ct #ttggagtc    420 agtgctcagc ttgatcgaac agtgggcagg aggaatactt tcattggaac tc #cctactgg    480 atggcaccag aagttattgc ctgtgatgaa aacccagatg ccacatatga tt #tcaagagt    540 gacttgtggt ctttgggtat caccgccatt gaaatggcag aaggtgctcc cc #ctctctgt    600 gacatgcacc ccatgagagc tctcttcctc atcccccgga atccagcgcc tc #ggctgaag    660 tctaagaagt ggtcaaaaaa attccagtca tttattgaga gctgcttggt aa #agaatcac    720 agccagcgac cagcaacaga acaattgatg aagcatccat ttatacgaga cc #aacctaat    780 gagcgacagg tccgcattca actcaaggac catattgata gaacaaagaa ga #agcgagga    840 gaaaaagatg agacagagta tgagtacagt ggaagtgagg aagaagagga gg #agaatgac    900 tcaggagagc ccagctccat cctgaatctg ccaagggagt cgacgctgcg ga #gggacttt    960 ctgaggctgc agctggccaa caaggagcgt tctgaggccc tacggaggca gc #agctggag   1020 cagcagcagc gggagaatga ggagcacaag cggcagctgc tggccgagcg tc #agaagcgc   1080 atcgaggagc agaaagagca gaggcggcgg ctggaggagc aacaaaggcg ag #agaaggag   1140 ctgcggaagc agcaggagag ggagcagcgc cggcactatg aggagcagat gc #gccgggag   1200 gaggagagga ggcgtgcgga gcatgaacag gaatataagc gcaaacaatt gg #aagaacag   1260 agacaagcag aaagactgca gaggcagcta aagcaagaaa gagactactt ag #tttccctt   1320 cagcatcagc ggcaggagca gaggcctgtg gagaagaagc cactgtacca tt #acaaagaa   1380 ggaatgagtc ctagtgagaa gccagcatgg gccaaggagg tagaagaacg gt #caaggctc   1440 aaccggcaaa gttcccctgc catgcctcac aaggttgcca acaggatatc tg #accccaac   1500 ctgcccccaa ggtcggagtc cttcagcatt agtggagttc agcctgctcg aa #cacccccc   1560 atgctcagac cagtcgatcc ccagatccca catctggtag ctgtaaaatc cc #agggacct   1620 gccttgaccg cctcccagtc agtgcacgag cagcccacaa agggcctctc tg #ggtttcag   1680 gaggctctga acgtgacctc ccaccgcgtg gagatgccac gccagaactc ag #atcccacc   1740 tcggaaaatc ctcctctccc cactcgcatt gaaaagtttg accgaagctc tt #ggttacga   1800 caggaagaag acattccacc aaaggtgcct caaagaacaa cttctatatc cc #cagcatta   1860 gccagaaaga attctcctgg gaatggtagt gctctgggac ccagactagg at #ctcaaccc   1920 atcagagcaa gcaaccctga tctccggaga actgagccca tcttggagag cc #ccttgcag   1980 aggaccagca gtggcagttc ctccagctcc agcaccccta gctcccagcc ca #gctcccaa   2040 ggaggctccc agcctggatc acaagcagga tccagtgaac gcaccagagt tc #gagccaac   2100 agtaagtcag aaggatcacc tgtgctcccc catgagcctg ccaaggtgaa ac #cagaagaa   2160 tccagggaca ttacccggcc cagtcgacca gctagctaca aaaaagctat ag #atgaggat   2220 ctgacggcat tagccaaaga actaagagaa ctccggattg aagaaacaaa cc #gcccaatg   2280 aagaaggtga ctgattactc ctcctccagt gaggagtcag aaagtagcga gg #aagaggag   2340 gaagatggag agagcgagac ccatgatggg acagtggctg tcagcgacat ac #ccagactg   2400 ataccaacag gagctccagg cagcaacgag cagtacaatg tgggaatggt gg #ggacgcat   2460 gggctggaga cctctcatgc ggacagtttc agcggcagta tttcaagaga ag #gaaccttg   2520 atgattagag agacgtctgg agagaagaag cgatctggcc acagtgacag ca #atggcttt   2580 gctggccaca tcaacctccc tgacctggtg cagcagagcc attctccagc tg #gaaccccg   2640 actgagggac tggggcgcgt ctcaacccat tcccaggaga tggactctgg ga #ctgaatat   2700 ggcatgggga gcagcaccaa agcctccttc accccctttg tggaccccag ag #tataccag   2760 acgtctccca ctgatgaaga tgaagaggat gaggaatcat cagccgcagc tc #tgtttact   2820 ggcgaacttc ttaggcaaga acaggccaaa ctcaatgaag caagaaagat tt #cggtggta   2880 aatgtaaacc caaccaacat tcggcctcat agcgacacac cagaaatcag aa #aatacaag   2940 aaacgattca actcagaaat actttgtgca gctctgtggg gtgtaaacct tc #tggtgggg   3000 actgaaaatg gcctgatgct tttggaccga agtgggcaag gcaaagtcta ta #atctgatc   3060 aaccggaggc gatttcagca gatggatgtg ctagagggac tgaatgtcct tg #tgacaatt   3120 tcaggaaaga agaataagct acgagtttac tatctttcat ggttaagaaa ca #gaatacta   3180 cataatgacc cagaagtaga aaagaaacaa ggctggatca ctgttgggga ct #tggaaggc   3240 tgtatacatt ataaagttgt taaatatgaa aggatcaaat ttttggtgat tg #ccttaaag   3300 aatgctgtgg aaatatatgc ttgggctcct aaaccgtatc ataaattcat gg #catttaag   3360 tcttttgcag atctccagca caagcctctg ctagttgatc tcacggtaga ag #aaggtcaa   3420 agattaaagg ttatttttgg ttcacacact ggtttccatg taattgatgt tg #attcagga   3480 aactcttatg atatctacac accatctcat attcagggca atatcactcc tc #atgctatt   3540 gtcatcttgc ctaaaacaga tggaatggaa atgcttgttt gctatgagga tg #agggggtg   3600 tatgtaaaca cctatggccg gataactaag gatgtggtgc tccaatgggg ag #aaatgccc   3660 acgtctgtgg cctacattca ttccaatcag ataatgggct ggggcgagaa ag #ctattgag   3720 atccggtcag tggaaacagg acatttggat ggagtattta tgcataagcg ag #ctcaaagg   3780 ttaaagtttc tatgtgaaag aaatgataag gtattttttg catccgtgcg at #ctggagga   3840 agtagccaag tgtttttcat gaccctcaac agaaattcca tgatgaactg gt #aacagaag   3900 agcacttggc acttatcttc atggcgttat ttctaattta aaagaacata ac #tcatgtgg   3960 acttatgcca gtctagaggc agaatcagaa ggcttggttg aacatatcgc tt #tccctttt   4020 tcctctccct ccgcccctcc cagtacagtc catct        #                   #     4055 <210> SEQ ID NO 11 <211> LENGTH: 4133 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 11 gcatttgggg aggtgtatga gggtcggcat gtcaagacgg ggcagctggc tg #ccatcaag     60 gtcatggatg tcacggagga cgaggaggaa gagatcaaac aggagatcaa ca #tgctgaaa    120 aagtactctc accaccgcaa catcgccacc tactacggag ccttcatcaa ga #agagcccc    180 ccgggaaacg atgaccagct ctggctggtg atggagttct gtggtgctgg tt #cagtgact    240 gacctggtaa agaacacaaa aggcaacgcc ctgaaggagg actgtatcgc ct #atatctgc    300 agggagatcc tcaggggtct ggcccatctc catgcccaca aggtgatcca tc #gagacatc    360 aaggggcaga atgtgctgct gacagagaat gctgaggtca agctagtgga tt #ttggggtg    420 agtgctcagc tggaccgcac cgtgggcaga cggaacactt tcattgggac tc #cctactgg    480 atggctccag aggtcatcgc ctgtgatgag aaccctgatg ccacctatga tt #acaggagt    540 gatatttggt ctctaggaat cacagccatc gagatggcag agggagcccc cc #ctctgtgt    600 gacatgcacc ccatgcgagc cctcttcctc attcctcgga accctccgcc ca #ggctcaag    660 tccaagaagt ggtctaagaa gttcattgac ttcattgaca catgtctcat ca #agacttac    720 ctgagccgcc cacccacgga gcagctactg aagtttccct tcatccggga cc #agcccacg    780 gagcggcagg tccgcatcca gcttaaggac cacattgacc gatcccggaa ga #agcggggt    840 gagaaagagg agacagaata tgagtacagc ggcagcgagg aggaagatga ca #gccatgga    900 gaggaaggag agccaagctc catcatgaac gtgcctggag agtcgactct ac #gccgggag    960 tttctccggc tccagcagga aaataagagc aactcagagg ctttaaaaca gc #agcagcag   1020 ctgcagcagc agcagcagcg agaccccgag gcacacatca aacacctgct gc #accagcgg   1080 cagcggcgca tagaggagca gaaggaggag cggcgccgcg tggaggagca ac #agcggcgg   1140 gagcgggagc agcggaagct gcaggagaag gagcagcagc ggcggctgga gg #acatgcag   1200 gctctgcggc gggaggagga gcggcggcag gcggagcgcg agcaggaata ta #ttcgtcac   1260 aggctagagg aggagcagcg acagctcgag atccttcagc aacagctgct cc #aggaacag   1320 gccctgctgc tggaatacaa gcggaagcag ctggaggagc agcggcagtc ag #aacgtctc   1380 cagaggcagc tgcagcagga gcatgcctac ctcaagtccc tgcagcagca gc #aacagcag   1440 cagcagcttc agaaacaaca gcagcagcag ctcctgcctg gggacaggaa gc #ccctgtac   1500 cattatggtc ggggcatgaa tcccgctgac aaaccagcct gggcccgaga gg #tagaagag   1560 agaacaagga tgaacaagca gcagaactct cccttggcca agagcaagcc ag #gcagcacg   1620 gggcctgagc cccccatccc ccaggcctcc ccagggcccc caggacccct tt #cccagact   1680 cctcctatgc agaggccggt ggagccccag gagggaccgc acaagagcct gg #tggcacac   1740 cgggtcccac tgaagccata tgcagcacct gtaccccgat cccagtccct gc #aggaccag   1800 cccacccgaa acctggctgc cttcccagcc tcccatgacc ccgaccctgc ca #tccccgca   1860 cccactgcca cgcccagtgc ccgaggagct gtcatccgcc agaattcaga cc #ccacctct   1920 gaaggacctg gccccagccc gaatccccca gcctgggtcc gcccagataa cg #aggcccca   1980 cccaaggtgc ctcagaggac ctcatctatc gccactgccc ttaacaccag tg #gggccgga   2040 gggtcccggc cagcccaggc agtccgtgcc agacctcgca gcaactccgc ct #ggcaaatc   2100 tatctgcaaa ggcgggcaga gcggggcacc ccaaagcctc cagggccccc tg #ctcagccc   2160 cctggcccgc ccaacgcctc tagtaacccc gacctcagga ggagcgaccc tg #gctgggaa   2220 cgctcggaca gcgtccttcc agcctctcac gggcacctcc cccaggctgg ct #cactggag   2280 cggaaccgcg tgggagtctc ctccaaaccg gacagctccc ctgtgctctc cc #ctgggaat   2340 aaagccaagc ccgacgacca ccgctcacgg ccaggccggc ccgcagactt tg #tgttgctg   2400 aaagagcgga ctctggacga ggcccctcgg cctcccaaga aggccatgga ct #actcgtcg   2460 tccagcgagg aggtggaaag cagtgaggac gacgaggagg aaggcgaagg cg #ggccagca   2520 gaggggagca gagatacccc tgggggccgc gatggggata cagacagcgt ca #gcaccatg   2580 gtggtccacg acgtcgagga gatcaccggg acccagcccc catacggggg cg #gcaccatg   2640 gtggtccagc gcacccctga agaggagcgg aacctgctgc atgctgacag ca #atgggtac   2700 acaaacctgc ctgacgtggt ccagcccagc cactcaccca ccgagaacag ca #aaggccaa   2760 agcccaccct cgaaggatgg gagtggtgac taccagtctc gtgggctggt aa #aggcccct   2820 ggcaagagct cgttcacgat gtttgtggat ctagggatct accagcctgg ag #gcagtggg   2880 gacagcatcc ccatcacagc cctagtgggt ggagagggca ctcggctcga cc #agctgcag   2940 tacgacgtga ggaagggttc tgtggtcaac gtgaatccca ccaacacccg gg #cccacagt   3000 gagacccctg agatccggaa gtacaagaag cgattcaact ccgagatcct ct #gtgcagcc   3060 ctttgggggg tcaacctgct ggtgggcacg gagaacgggc tgatgttgct gg #accgaagt   3120 gggcagggca aggtgtatgg actcattggg cggcgacgct tccagcagat gg #atgtgctg   3180 gaggggctca acctgctcat caccatctca gggaaaagga acaaactgcg gg #tgtattac   3240 ttgtcctggc tccggaacaa gattctgcac aatgacccag aagtggagaa ga #agcagggc   3300 tggaccaccg tgggggacat ggagggctgc gggcactacc gtgttgtgaa at #acgagcgg   3360 attaagttcc tggtcatcgc cctcaagagc tccgtggagg tgtatgcctg gg #cccccaaa   3420 ccctaccaca aattcatggc cttcaagtcc tttgccgacc tcccccaccg cc #ctctgctg   3480 gtcgacctga cagtagagga ggggcagcgg ctcaaggtca tctatggctc ca #gtgctggc   3540 ttccatgctg tggatgtcga ctcggggaac agctatgaca tctacatccc tg #tgcacatc   3600 cagagccaga tcacgcccca tgccatcatc ttcctcccca acaccgacgg ca #tggagatg   3660 ctgctgtgct acgaggacga gggtgtctac gtcaacacgt acgggcgcat ca #ttaaggat   3720 gtggtgctgc agtgggggga gatgcctact tctgtggcct acatctgctc ca #accagata   3780 atgggctggg gtgagaaagc cattgagatc cgctctgtgg agacgggcca cc #tcgacggg   3840 gtcttcatgc acaaacgagc tcagaggctc aagttcctgt gtgagcggaa tg #acaaggtg   3900 ttttttgcct cagtccgctc tgggggcagc agccaagttt acttcatgac tc #tgaaccgt   3960 aaccgcatca tgaactggtg acggggccct gggctggggc tgtcccacac tg #gacccagc   4020 tctccccctg cagccaggct tcccgggccg cccctctttc ccctccctgg gc #ttttgctt   4080 ttactggttt gatttcactg gagcctgctg ggaacgtgac ctctgacccc tg #a          4133 <210> SEQ ID NO 12 <211> LENGTH: 1459 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 12 caatgttaac ccactctatg tctctcctgc atgtaaaaaa ccactaatcc ac #atgtatga     60 aaaggagttc acttctgaga tctgctgtgg ttctttgtgg ggagtcaatt tg #ctgttggg    120 aacccgatct aatctatatc tgatggacag aagtggaaag gctgacatta ct #aaacttat    180 aaggcgaaga ccattccgcc agattcaagt cttagagcca ctcaatttgc tg #attaccat    240 ctcaggtcat aagaacagac ttcgggtgta tcatctgacc tggttgagga ac #aagatttt    300 gaataatgat ccagaaagta aaagaaggca agaagaaatg ctgaagacag ag #gaagcctg    360 caaagctatt gataagttaa caggctgtga acacttcagt gtcctccaac at #gaagaaac    420 aacatatatt gcaattgctt tgaaatcatc aattcacctt tatgcatggg ca #ccaaagtc    480 ctttgatgaa agcactgcta ttaaagtatt tccaacactt gatcataagc ca #gtgacagt    540 tgacctggct attggttctg aaaaaagact aaagattttc ttcagctcag ca #gatggata    600 tcacctcatc gatgcagaat ctgaggttat gtctgatgtg accctgccaa ag #aatcccct    660 ggaaatcatt ataccacaga atatcatcat tttacctgat tgcttgggaa tt #ggcatgat    720 gctcaccttc aatgctgaag ccctctctgt ggaagcaaat gaacaactct tc #aagaagat    780 ccttgaaatg tggaaagaca taccatcttc tatagctttt gaatgtacac ag #cgaaccac    840 aggatggggc caaaaggcca ttgaagtgcg ctctttgcaa tccagggttc tg #gaaagtga    900 gctgaagcgc aggtcaatta agaagctgag attcctgtgc acccggggtg ac #aagctgtt    960 ctttacctct accctgcgca atcaccacag ccgggtttac ttcatgacac tt #ggaaaact   1020 tgaagagctc caaagcaatt atgatgtcta aaagtttcca gtgatttatt ac #cacattat   1080 aaacatcatg tataggcagt ctgcatcttc agatttcaga gattaaatga gt #attcagtt   1140 ttatttttag taaagattaa atccaaaact ttacttttaa tgtagcacag aa #tagtttta   1200 atgagaaatg cagctttatg tataaaatta actatagcaa gctctaggta ct #ccaatggt   1260 gtacaatgtc ttttgcacaa actttgtaac ttttgttact gtgaattcaa ac #attactct   1320 ttggacagtt tggacagtat ctgtattcag attttacaac atggagtaaa ga #aacctgtt   1380 atgaattaga ttacaagcag ccttcaaaag aattggcact gggataagat tt #ttcagaaa   1440 agaaaaacat cggcaaact              #                   #                 145 #9 <210> SEQ ID NO 13 <211> LENGTH: 1239 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 13 Met Ala Asn Asp Ser Pro Ala Lys Ser Leu Va #l Asp Ile Asp Leu Ser   1               5  #                 10  #                 15 Ser Leu Arg Asp Pro Ala Gly Ile Phe Glu Le #u Val Glu Val Val Gly              20      #             25      #             30 Asn Gly Thr Tyr Gly Gln Val Tyr Lys Gly Ar #g His Val Lys Thr Gly          35          #         40          #         45 Gln Leu Ala Ala Ile Lys Val Met Asp Val Th #r Glu Asp Glu Glu Glu      50              #     55              #     60 Glu Ile Lys Leu Glu Ile Asn Met Leu Lys Ly #s Tyr Ser His His Arg  65                  # 70                  # 75                  # 80 Asn Ile Ala Thr Tyr Tyr Gly Ala Phe Ile Ly #s Lys Ser Pro Pro Gly                  85  #                 90  #                 95 His Asp Asp Gln Leu Trp Leu Val Met Glu Ph #e Cys Gly Ala Gly Ser             100       #           105       #           110 Ile Thr Asp Leu Val Lys Asn Thr Lys Gly As #n Thr Leu Lys Glu Asp         115           #       120           #       125 Trp Ile Ala Tyr Ile Ser Arg Glu Ile Leu Ar #g Gly Leu Ala His Leu     130               #   135               #   140 His Ile His His Val Ile His Arg Asp Ile Ly #s Gly Gln Asn Val Leu 145                 1 #50                 1 #55                 1 #60 Leu Thr Glu Asn Ala Glu Val Lys Leu Val As #p Phe Gly Val Ser Ala                 165   #               170   #               175 Gln Leu Asp Arg Thr Val Gly Arg Arg Asn Th #r Phe Ile Gly Thr Pro             180       #           185       #           190 Tyr Trp Met Ala Pro Glu Val Ile Ala Cys As #p Glu Asn Pro Asp Ala         195           #       200           #       205 Thr Tyr Asp Tyr Arg Ser Asp Leu Trp Ser Cy #s Gly Ile Thr Ala Ile     210               #   215               #   220 Glu Met Ala Glu Gly Ala Pro Pro Leu Cys As #p Met His Pro Met Arg 225                 2 #30                 2 #35                 2 #40 Ala Leu Phe Leu Ile Pro Arg Asn Pro Pro Pr #o Arg Leu Lys Ser Lys                 245   #               250   #               255 Lys Trp Ser Lys Lys Phe Phe Ser Phe Ile Gl #u Gly Cys Leu Val Lys             260       #           265       #           270 Asn Tyr Met Gln Arg Pro Ser Thr Glu Gln Le #u Leu Lys His Pro Phe         275           #       280           #       285 Ile Arg Asp Gln Pro Asn Glu Arg Gln Val Ar #g Ile Gln Leu Lys Asp     290               #   295               #   300 His Ile Asp Arg Thr Arg Lys Lys Arg Gly Gl #u Lys Asp Glu Thr Glu 305                 3 #10                 3 #15                 3 #20 Tyr Glu Tyr Ser Gly Ser Glu Glu Glu Glu Gl #u Glu Val Pro Glu Gln                 325   #               330   #               335 Glu Gly Glu Pro Ser Ser Ile Val Asn Val Pr #o Gly Glu Ser Thr Leu             340       #           345       #           350 Arg Arg Asp Phe Leu Arg Leu Gln Gln Glu As #n Lys Glu Arg Ser Glu         355           #       360           #       365 Ala Leu Arg Arg Gln Gln Leu Leu Gln Glu Gl #n Gln Leu Arg Glu Gln     370               #   375               #   380 Glu Glu Tyr Lys Arg Gln Leu Leu Ala Glu Ar #g Gln Lys Arg Ile Glu 385                 3 #90                 3 #95                 4 #00 Gln Gln Lys Glu Gln Arg Arg Arg Leu Glu Gl #u Gln Gln Arg Arg Glu                 405   #               410   #               415 Arg Glu Ala Arg Arg Gln Gln Glu Arg Glu Gl #n Arg Arg Arg Glu Gln             420       #           425       #           430 Glu Glu Lys Arg Arg Leu Glu Glu Leu Glu Ar #g Arg Arg Lys Glu Glu         435           #       440           #       445 Glu Glu Arg Arg Arg Ala Glu Glu Glu Lys Ar #g Arg Val Glu Arg Glu     450               #   455               #   460 Gln Glu Tyr Ile Arg Arg Gln Leu Glu Glu Gl #u Gln Arg His Leu Glu 465                 4 #70                 4 #75                 4 #80 Val Leu Gln Gln Gln Leu Leu Gln Glu Gln Al #a Met Leu Leu Glu Cys                 485   #               490   #               495 Arg Trp Arg Glu Met Glu Glu His Arg Gln Al #a Glu Arg Leu Gln Arg             500       #           505       #           510 Gln Leu Gln Gln Glu Gln Ala Tyr Leu Leu Se #r Leu Gln His Asp His         515           #       520           #       525 Arg Arg Pro His Pro Gln His Ser Gln Gln Pr #o Pro Pro Pro Gln Gln     530               #   535               #   540 Glu Arg Ser Lys Pro Ser Phe His Ala Pro Gl #u Pro Lys Ala His Tyr 545                 5 #50                 5 #55                 5 #60 Glu Pro Ala Asp Arg Ala Arg Glu Val Glu As #p Arg Phe Arg Lys Thr                 565   #               570   #               575 Asn His Ser Ser Pro Glu Ala Gln Ser Lys Gl #n Thr Gly Arg Val Leu             580       #           585       #           590 Glu Pro Pro Val Pro Ser Arg Ser Glu Ser Ph #e Ser Asn Gly Asn Ser         595           #       600           #       605 Glu Ser Val His Pro Ala Leu Gln Arg Pro Al #a Glu Pro Gln Val Pro     610               #   615               #   620 Val Arg Thr Thr Ser Arg Ser Pro Val Leu Se #r Arg Arg Asp Ser Pro 625                 6 #30                 6 #35                 6 #40 Leu Gln Gly Ser Gly Gln Gln Asn Ser Gln Al #a Gly Gln Arg Asn Ser                 645   #               650   #               655 Thr Ser Ile Glu Pro Arg Leu Leu Trp Glu Ar #g Val Glu Lys Leu Val             660       #           665       #           670 Pro Arg Pro Gly Ser Gly Ser Ser Ser Gly Se #r Ser Asn Ser Gly Ser         675           #       680           #       685 Gln Pro Gly Ser His Pro Gly Ser Gln Ser Gl #y Ser Gly Glu Arg Phe     690               #   695               #   700 Arg Val Arg Ser Ser Ser Lys Ser Glu Gly Se #r Pro Ser Gln Arg Leu 705                 7 #10                 7 #15                 7 #20 Glu Asn Ala Val Lys Lys Pro Glu Asp Lys Ly #s Glu Val Phe Arg Pro                 725   #               730   #               735 Leu Lys Pro Ala Asp Leu Thr Ala Leu Ala Ly #s Glu Leu Arg Ala Val             740       #           745       #           750 Glu Asp Val Arg Pro Pro His Lys Val Thr As #p Tyr Ser Ser Ser Ser         755           #       760           #       765 Glu Glu Ser Gly Thr Thr Asp Glu Glu Asp As #p Asp Val Glu Gln Glu     770               #   775               #   780 Gly Ala Asp Glu Ser Thr Ser Gly Pro Glu As #p Thr Arg Ala Ala Ser 785                 7 #90                 7 #95                 8 #00 Ser Leu Asn Leu Ser Asn Gly Glu Thr Glu Se #r Val Lys Thr Met Ile                 805   #               810   #               815 Val His Asp Asp Val Glu Ser Glu Pro Ala Me #t Thr Pro Ser Lys Glu             820       #           825       #           830 Gly Thr Leu Ile Val Arg Arg Thr Gln Ser Al #a Ser Ser Thr Leu Gln         835           #       840           #       845 Lys His Lys Ser Ser Ser Ser Phe Thr Pro Ph #e Ile Asp Pro Arg Leu     850               #   855               #   860 Leu Gln Ile Ser Pro Ser Ser Gly Thr Thr Va #l Thr Ser Val Val Gly 865                 8 #70                 8 #75                 8 #80 Phe Ser Cys Asp Gly Met Arg Pro Glu Ala Il #e Arg Gln Asp Pro Thr                 885   #               890   #               895 Arg Lys Gly Ser Val Val Asn Val Asn Pro Th #r Asn Thr Arg Pro Gln             900       #           905       #           910 Ser Asp Thr Pro Glu Ile Arg Lys Tyr Lys Ly #s Arg Phe Asn Ser Glu         915           #       920           #       925 Ile Leu Cys Ala Ala Leu Trp Gly Val Asn Le #u Leu Val Gly Thr Glu     930               #   935               #   940 Ser Gly Leu Met Leu Leu Asp Arg Ser Gly Gl #n Gly Lys Val Tyr Pro 945                 9 #50                 9 #55                 9 #60 Leu Ile Asn Arg Arg Arg Phe Gln Gln Met As #p Val Leu Glu Gly Leu                 965   #               970   #               975 Asn Val Leu Val Thr Ile Ser Gly Lys Lys As #p Lys Leu Arg Val Tyr             980       #           985       #           990 Tyr Leu Ser Trp Leu Arg Asn Lys Ile Leu Hi #s Asn Asp Pro Glu Val         995           #      1000            #     1005 Glu Lys Lys Gln Gly Trp Thr Thr Val Gly As #p Leu Glu Gly Cys Val    1010               #  1015                # 1020 His Tyr Lys Val Val Lys Tyr Glu Arg Ile Ly #s Phe Leu Val Ile Ala 1025               1030  #               1035   #              1040 Leu Lys Ser Ser Val Glu Val Tyr Ala Trp Al #a Pro Lys Pro Tyr His                1045   #              1050    #             1055 Lys Phe Met Ala Phe Lys Ser Phe Gly Glu Le #u Val His Lys Pro Leu            1060       #          1065        #         1070 Leu Val Asp Leu Thr Val Glu Glu Gly Gln Ar #g Leu Lys Val Ile Tyr        1075           #      1080            #     1085 Gly Ser Cys Ala Gly Phe His Ala Val Asp Va #l Asp Ser Gly Ser Val    1090               #  1095                # 1100 Tyr Asp Ile Tyr Leu Pro Thr His Ile Gln Cy #s Ser Ile Lys Pro His 1105               1110  #               1115   #              1120 Ala Ile Ile Ile Leu Pro Asn Thr Asp Gly Me #t Glu Leu Leu Val Cys                1125   #              1130    #             1135 Tyr Glu Asp Glu Gly Val Tyr Val Asn Thr Ty #r Gly Arg Ile Thr Lys            1140       #          1145        #         1150 Asp Val Val Leu Gln Trp Gly Glu Met Pro Th #r Ser Val Ala Tyr Ile        1155           #      1160            #     1165 Arg Ser Asn Gln Thr Met Gly Trp Gly Glu Ly #s Ala Ile Glu Ile Arg    1170               #  1175                # 1180 Ser Val Glu Thr Gly His Leu Asp Gly Val Ph #e Met His Lys Arg Ala 1185               1190  #               1195   #              1200 Gln Arg Leu Lys Phe Leu Cys Glu Arg Asn As #p Lys Val Phe Phe Ala                1205   #              1210    #             1215 Ser Val Arg Ser Gly Gly Ser Ser Gln Val Ty #r Phe Met Thr Leu Gly            1220       #          1225        #         1230 Arg Thr Ser Leu Leu Ser Trp        1235 <210> SEQ ID NO 14 <211> LENGTH: 1297 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 14 Ala Phe Gly Glu Val Tyr Glu Gly Arg His Va #l Lys Thr Gly Gln Leu   1               5  #                 10  #                 15 Ala Ala Ile Lys Val Met Asp Val Thr Gly As #p Glu Glu Glu Glu Ile              20      #             25      #             30 Lys Gln Glu Ile Asn Met Leu Lys Lys Tyr Se #r His His Arg Asn Ile          35          #         40          #         45 Ala Thr Tyr Tyr Gly Ala Phe Ile Lys Lys As #n Pro Pro Gly Met Asp      50              #     55              #     60 Asp Gln Leu Trp Leu Val Met Glu Phe Cys Gl #y Ala Gly Ser Val Thr  65                  # 70                  # 75                  # 80 Asp Leu Ile Lys Asn Thr Lys Gly Asn Thr Le #u Lys Glu Glu Trp Ile                  85  #                 90  #                 95 Ala Tyr Ile Cys Arg Glu Ile Leu Arg Gly Le #u Ser His Leu His Gln             100       #           105       #           110 His Lys Val Ile His Arg Asp Ile Lys Gly Gl #n Asn Val Leu Leu Thr         115           #       120           #       125 Glu Asn Ala Glu Val Lys Leu Val Asp Phe Gl #y Val Ser Ala Gln Leu     130               #   135               #   140 Asp Arg Thr Val Gly Arg Arg Asn Thr Phe Il #e Gly Thr Pro Tyr Trp 145                 1 #50                 1 #55                 1 #60 Met Ala Pro Glu Val Ile Ala Cys Asp Glu As #n Pro Asp Ala Thr Tyr                 165   #               170   #               175 Asp Phe Lys Ser Asp Leu Trp Ser Leu Gly Il #e Thr Ala Ile Glu Met             180       #           185       #           190 Ala Glu Gly Ala Pro Pro Leu Cys Asp Met Hi #s Pro Met Arg Ala Leu         195           #       200           #       205 Phe Leu Ile Pro Arg Asn Pro Ala Pro Arg Le #u Lys Ser Lys Lys Trp     210               #   215               #   220 Ser Lys Lys Phe Gln Ser Phe Ile Glu Ser Cy #s Leu Val Lys Asn His 225                 2 #30                 2 #35                 2 #40 Ser Gln Arg Pro Ala Thr Glu Gln Leu Met Ly #s His Pro Phe Ile Arg                 245   #               250   #               255 Asp Gln Pro Asn Glu Arg Gln Val Arg Ile Gl #n Leu Lys Asp His Ile             260       #           265       #           270 Asp Arg Thr Lys Lys Lys Arg Gly Glu Lys As #p Glu Thr Glu Tyr Glu         275           #       280           #       285 Tyr Ser Gly Ser Glu Glu Glu Glu Glu Glu As #n Asp Ser Gly Glu Pro     290               #   295               #   300 Ser Ser Ile Leu Asn Leu Pro Arg Glu Ser Th #r Leu Arg Arg Asp Phe 305                 3 #10                 3 #15                 3 #20 Leu Arg Leu Gln Leu Ala Asn Lys Glu Arg Se #r Glu Ala Leu Arg Arg                 325   #               330   #               335 Gln Gln Leu Glu Gln Gln Gln Arg Glu Asn Gl #u Glu His Lys Arg Gln             340       #           345       #           350 Leu Leu Ala Glu Arg Gln Lys Arg Ile Glu Gl #u Gln Lys Glu Gln Arg         355           #       360           #       365 Arg Arg Leu Glu Glu Gln Gln Arg Arg Glu Ly #s Glu Leu Arg Lys Gln     370               #   375               #   380 Gln Glu Arg Glu Gln Arg Arg His Tyr Glu Gl #u Gln Met Arg Arg Glu 385                 3 #90                 3 #95                 4 #00 Glu Glu Arg Arg Arg Ala Glu His Glu Gln Gl #u Tyr Lys Arg Lys Gln                 405   #               410   #               415 Leu Glu Glu Gln Arg Gln Ala Glu Arg Leu Gl #n Arg Gln Leu Lys Gln             420       #           425       #           430 Glu Arg Asp Tyr Leu Val Ser Leu Gln His Gl #n Arg Gln Glu Gln Arg         435           #       440           #       445 Pro Val Glu Lys Lys Pro Leu Tyr His Tyr Ly #s Glu Gly Met Ser Pro     450               #   455               #   460 Ser Glu Lys Pro Ala Trp Ala Lys Glu Val Gl #u Glu Arg Ser Arg Leu 465                 4 #70                 4 #75                 4 #80 Asn Arg Gln Ser Ser Pro Ala Met Pro His Ly #s Val Ala Asn Arg Ile                 485   #               490   #               495 Ser Asp Pro Asn Leu Pro Pro Arg Ser Glu Se #r Phe Ser Ile Ser Gly             500       #           505       #           510 Val Gln Pro Ala Arg Thr Pro Pro Met Leu Ar #g Pro Val Asp Pro Gln         515           #       520           #       525 Ile Pro His Leu Val Ala Val Lys Ser Gln Gl #y Pro Ala Leu Thr Ala     530               #   535               #   540 Ser Gln Ser Val His Glu Gln Pro Thr Lys Gl #y Leu Ser Gly Phe Gln 545                 5 #50                 5 #55                 5 #60 Glu Ala Leu Asn Val Thr Ser His Arg Val Gl #u Met Pro Arg Gln Asn                 565   #               570   #               575 Ser Asp Pro Thr Ser Glu Asn Pro Pro Leu Pr #o Thr Arg Ile Glu Lys             580       #           585       #           590 Phe Asp Arg Ser Ser Trp Leu Arg Gln Glu Gl #u Asp Ile Pro Pro Lys         595           #       600           #       605 Val Pro Gln Arg Thr Thr Ser Ile Ser Pro Al #a Leu Ala Arg Lys Asn     610               #   615               #   620 Ser Pro Gly Asn Gly Ser Ala Leu Gly Pro Ar #g Leu Gly Ser Gln Pro 625                 6 #30                 6 #35                 6 #40 Ile Arg Ala Ser Asn Pro Asp Leu Arg Arg Th #r Glu Pro Ile Leu Glu                 645   #               650   #               655 Ser Pro Leu Gln Arg Thr Ser Ser Gly Ser Se #r Ser Ser Ser Ser Thr             660       #           665       #           670 Pro Ser Ser Gln Pro Ser Ser Gln Gly Gly Se #r Gln Pro Gly Ser Gln         675           #       680           #       685 Ala Gly Ser Ser Glu Arg Thr Arg Val Arg Al #a Asn Ser Lys Ser Glu     690               #   695               #   700 Gly Ser Pro Val Leu Pro His Glu Pro Ala Ly #s Val Lys Pro Glu Glu 705                 7 #10                 7 #15                 7 #20 Ser Arg Asp Ile Thr Arg Pro Ser Arg Pro Al #a Ser Tyr Lys Lys Ala                 725   #               730   #               735 Ile Asp Glu Asp Leu Thr Ala Leu Ala Lys Gl #u Leu Arg Glu Leu Arg             740       #           745       #           750 Ile Glu Glu Thr Asn Arg Pro Met Lys Lys Va #l Thr Asp Tyr Ser Ser         755           #       760           #       765 Ser Ser Glu Glu Ser Glu Ser Ser Glu Glu Gl #u Glu Glu Asp Gly Glu     770               #   775               #   780 Ser Glu Thr His Asp Gly Thr Val Ala Val Se #r Asp Ile Pro Arg Leu 785                 7 #90                 7 #95                 8 #00 Ile Pro Thr Gly Ala Pro Gly Ser Asn Glu Gl #n Tyr Asn Val Gly Met                 805   #               810   #               815 Val Gly Thr His Gly Leu Glu Thr Ser His Al #a Asp Ser Phe Ser Gly             820       #           825       #           830 Ser Ile Ser Arg Glu Gly Thr Leu Met Ile Ar #g Glu Thr Ser Gly Glu         835           #       840           #       845 Lys Lys Arg Ser Gly His Ser Asp Ser Asn Gl #y Phe Ala Gly His Ile     850               #   855               #   860 Asn Leu Pro Asp Leu Val Gln Gln Ser His Se #r Pro Ala Gly Thr Pro 865                 8 #70                 8 #75                 8 #80 Thr Glu Gly Leu Gly Arg Val Ser Thr His Se #r Gln Glu Met Asp Ser                 885   #               890   #               895 Gly Thr Glu Tyr Gly Met Gly Ser Ser Thr Ly #s Ala Ser Phe Thr Pro             900       #           905       #           910 Phe Val Asp Pro Arg Val Tyr Gln Thr Ser Pr #o Thr Asp Glu Asp Glu         915           #       920           #       925 Glu Asp Glu Glu Ser Ser Ala Ala Ala Leu Ph #e Thr Gly Glu Leu Leu     930               #   935               #   940 Arg Gln Glu Gln Ala Lys Leu Asn Glu Ala Ar #g Lys Ile Ser Val Val 945                 9 #50                 9 #55                 9 #60 Asn Val Asn Pro Thr Asn Ile Arg Pro His Se #r Asp Thr Pro Glu Ile                 965   #               970   #               975 Arg Lys Tyr Lys Lys Arg Phe Asn Ser Glu Il #e Leu Cys Ala Ala Leu             980       #           985       #           990 Trp Gly Val Asn Leu Leu Val Gly Thr Glu As #n Gly Leu Met Leu Leu         995           #      1000            #     1005 Asp Arg Ser Gly Gln Gly Lys Val Tyr Asn Le #u Ile Asn Arg Arg Arg    1010               #  1015                # 1020 Phe Gln Gln Met Asp Val Leu Glu Gly Leu As #n Val Leu Val Thr Ile 1025               1030  #               1035   #              1040 Ser Gly Lys Lys Asn Lys Leu Arg Val Tyr Ty #r Leu Ser Trp Leu Arg                1045   #              1050    #             1055 Asn Arg Ile Leu His Asn Asp Pro Glu Val Gl #u Lys Lys Gln Gly Trp            1060       #          1065        #         1070 Ile Thr Val Gly Asp Leu Glu Gly Cys Ile Hi #s Tyr Lys Val Val Lys        1075           #      1080            #     1085 Tyr Glu Arg Ile Lys Phe Leu Val Ile Ala Le #u Lys Asn Ala Val Glu    1090               #  1095                # 1100 Ile Tyr Ala Trp Ala Pro Lys Pro Tyr His Ly #s Phe Met Ala Phe Lys 1105               1110  #               1115   #              1120 Ser Phe Ala Asp Leu Gln His Lys Pro Leu Le #u Val Asp Leu Thr Val                1125   #              1130    #             1135 Glu Glu Gly Gln Arg Leu Lys Val Ile Phe Gl #y Ser His Thr Gly Phe            1140       #          1145        #         1150 His Val Ile Asp Val Asp Ser Gly Asn Ser Ty #r Asp Ile Tyr Thr Pro        1155           #      1160            #     1165 Ser His Ile Gln Gly Asn Ile Thr Pro His Al #a Ile Val Ile Leu Pro    1170               #  1175                # 1180 Lys Thr Asp Gly Met Glu Met Leu Val Cys Ty #r Glu Asp Glu Gly Val 1185               1190  #               1195   #              1200 Tyr Val Asn Thr Tyr Gly Arg Ile Thr Lys As #p Val Val Leu Gln Trp                1205   #              1210    #             1215 Gly Glu Met Pro Thr Ser Val Ala Tyr Ile Hi #s Ser Asn Gln Ile Met            1220       #          1225        #         1230 Gly Trp Gly Glu Lys Ala Ile Glu Ile Arg Se #r Val Glu Thr Gly His        1235           #      1240            #     1245 Leu Asp Gly Val Phe Met His Lys Arg Ala Gl #n Arg Leu Lys Phe Leu    1250               #  1255                # 1260 Cys Glu Arg Asn Asp Lys Val Phe Phe Ala Se #r Val Arg Ser Gly Gly 1265               1270  #               1275   #              1280 Ser Ser Gln Val Phe Phe Met Thr Leu Asn Ar #g Asn Ser Met Met Asn                1285   #              1290    #             1295 Trp <210> SEQ ID NO 15 <211> LENGTH: 1326 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 15 Ala Phe Gly Glu Val Tyr Glu Gly Arg His Va #l Lys Thr Gly Gln Leu   1               5  #                 10  #                 15 Ala Ala Ile Lys Val Met Asp Val Thr Glu As #p Glu Glu Glu Glu Ile              20      #             25      #             30 Lys Gln Glu Ile Asn Met Leu Lys Lys Tyr Se #r His His Arg Asn Ile          35          #         40          #         45 Ala Thr Tyr Tyr Gly Ala Phe Ile Lys Lys Se #r Pro Pro Gly Asn Asp      50              #     55              #     60 Asp Gln Leu Trp Leu Val Met Glu Phe Cys Gl #y Ala Gly Ser Val Thr  65                  # 70                  # 75                  # 80 Asp Leu Val Lys Asn Thr Lys Gly Asn Ala Le #u Lys Glu Asp Cys Ile                  85  #                 90  #                 95 Ala Tyr Ile Cys Arg Glu Ile Leu Arg Gly Le #u Ala His Leu His Ala             100       #           105       #           110 His Lys Val Ile His Arg Asp Ile Lys Gly Gl #n Asn Val Leu Leu Thr         115           #       120           #       125 Glu Asn Ala Glu Val Lys Leu Val Asp Phe Gl #y Val Ser Ala Gln Leu     130               #   135               #   140 Asp Arg Thr Val Gly Arg Arg Asn Thr Phe Il #e Gly Thr Pro Tyr Trp 145                 1 #50                 1 #55                 1 #60 Met Ala Pro Glu Val Ile Ala Cys Asp Glu As #n Pro Asp Ala Thr Tyr                 165   #               170   #               175 Asp Tyr Arg Ser Asp Ile Trp Ser Leu Gly Il #e Thr Ala Ile Glu Met             180       #           185       #           190 Ala Glu Gly Ala Pro Pro Leu Cys Asp Met Hi #s Pro Met Arg Ala Leu         195           #       200           #       205 Phe Leu Ile Pro Arg Asn Pro Pro Pro Arg Le #u Lys Ser Lys Lys Trp     210               #   215               #   220 Ser Lys Lys Phe Ile Asp Phe Ile Asp Thr Cy #s Leu Ile Lys Thr Tyr 225                 2 #30                 2 #35                 2 #40 Leu Ser Arg Pro Pro Thr Glu Gln Leu Leu Ly #s Phe Pro Phe Ile Arg                 245   #               250   #               255 Asp Gln Pro Thr Glu Arg Gln Val Arg Ile Gl #n Leu Lys Asp His Ile             260       #           265       #           270 Asp Arg Ser Arg Lys Lys Arg Gly Glu Lys Gl #u Glu Thr Glu Tyr Glu         275           #       280           #       285 Tyr Ser Gly Ser Glu Glu Glu Asp Asp Ser Hi #s Gly Glu Glu Gly Glu     290               #   295               #   300 Pro Ser Ser Ile Met Asn Val Pro Gly Glu Se #r Thr Leu Arg Arg Glu 305                 3 #10                 3 #15                 3 #20 Phe Leu Arg Leu Gln Gln Glu Asn Lys Ser As #n Ser Glu Ala Leu Lys                 325   #               330   #               335 Gln Gln Gln Gln Leu Gln Gln Gln Gln Gln Ar #g Asp Pro Glu Ala His             340       #           345       #           350 Ile Lys His Leu Leu His Gln Arg Gln Arg Ar #g Ile Glu Glu Gln Lys         355           #       360           #       365 Glu Glu Arg Arg Arg Val Glu Glu Gln Gln Ar #g Arg Glu Arg Glu Gln     370               #   375               #   380 Arg Lys Leu Gln Glu Lys Glu Gln Gln Arg Ar #g Leu Glu Asp Met Gln 385                 3 #90                 3 #95                 4 #00 Ala Leu Arg Arg Glu Glu Glu Arg Arg Gln Al #a Glu Arg Glu Gln Glu                 405   #               410   #               415 Tyr Ile Arg His Arg Leu Glu Glu Glu Gln Ar #g Gln Leu Glu Ile Leu             420       #           425       #           430 Gln Gln Gln Leu Leu Gln Glu Gln Ala Leu Le #u Leu Glu Tyr Lys Arg         435           #       440           #       445 Lys Gln Leu Glu Glu Gln Arg Gln Ser Glu Ar #g Leu Gln Arg Gln Leu     450               #   455               #   460 Gln Gln Glu His Ala Tyr Leu Lys Ser Leu Gl #n Gln Gln Gln Gln Gln 465                 4 #70                 4 #75                 4 #80 Gln Gln Leu Gln Lys Gln Gln Gln Gln Gln Le #u Leu Pro Gly Asp Arg                 485   #               490   #               495 Lys Pro Leu Tyr His Tyr Gly Arg Gly Met As #n Pro Ala Asp Lys Pro             500       #           505       #           510 Ala Trp Ala Arg Glu Val Glu Glu Arg Thr Ar #g Met Asn Lys Gln Gln         515           #       520           #       525 Asn Ser Pro Leu Ala Lys Ser Lys Pro Gly Se #r Thr Gly Pro Glu Pro     530               #   535               #   540 Pro Ile Pro Gln Ala Ser Pro Gly Pro Pro Gl #y Pro Leu Ser Gln Thr 545                 5 #50                 5 #55                 5 #60 Pro Pro Met Gln Arg Pro Val Glu Pro Gln Gl #u Gly Pro His Lys Ser                 565   #               570   #               575 Leu Val Ala His Arg Val Pro Leu Lys Pro Ty #r Ala Ala Pro Val Pro             580       #           585       #           590 Arg Ser Gln Ser Leu Gln Asp Gln Pro Thr Ar #g Asn Leu Ala Ala Phe         595           #       600           #       605 Pro Ala Ser His Asp Pro Asp Pro Ala Ile Pr #o Ala Pro Thr Ala Thr     610               #   615               #   620 Pro Ser Ala Arg Gly Ala Val Ile Arg Gln As #n Ser Asp Pro Thr Ser 625                 6 #30                 6 #35                 6 #40 Glu Gly Pro Gly Pro Ser Pro Asn Pro Pro Al #a Trp Val Arg Pro Asp                 645   #               650   #               655 Asn Glu Ala Pro Pro Lys Val Pro Gln Arg Th #r Ser Ser Ile Ala Thr             660       #           665       #           670 Ala Leu Asn Thr Ser Gly Ala Gly Gly Ser Ar #g Pro Ala Gln Ala Val         675           #       680           #       685 Arg Ala Arg Pro Arg Ser Asn Ser Ala Trp Gl #n Ile Tyr Leu Gln Arg     690               #   695               #   700 Arg Ala Glu Arg Gly Thr Pro Lys Pro Pro Gl #y Pro Pro Ala Gln Pro 705                 7 #10                 7 #15                 7 #20 Pro Gly Pro Pro Asn Ala Ser Ser Asn Pro As #p Leu Arg Arg Ser Asp                 725   #               730   #               735 Pro Gly Trp Glu Arg Ser Asp Ser Val Leu Pr #o Ala Ser His Gly His             740       #           745       #           750 Leu Pro Gln Ala Gly Ser Leu Glu Arg Asn Ar #g Val Gly Val Ser Ser         755           #       760           #       765 Lys Pro Asp Ser Ser Pro Val Leu Ser Pro Gl #y Asn Lys Ala Lys Pro     770               #   775               #   780 Asp Asp His Arg Ser Arg Pro Gly Arg Pro Al #a Asp Phe Val Leu Leu 785                 7 #90                 7 #95                 8 #00 Lys Glu Arg Thr Leu Asp Glu Ala Pro Arg Pr #o Pro Lys Lys Ala Met                 805   #               810   #               815 Asp Tyr Ser Ser Ser Ser Glu Glu Val Glu Se #r Ser Glu Asp Asp Glu             820       #           825       #           830 Glu Glu Gly Glu Gly Gly Pro Ala Glu Gly Se #r Arg Asp Thr Pro Gly         835           #       840           #       845 Gly Arg Asp Gly Asp Thr Asp Ser Val Ser Th #r Met Val Val His Asp     850               #   855               #   860 Val Glu Glu Ile Thr Gly Thr Gln Pro Pro Ty #r Gly Gly Gly Thr Met 865                 8 #70                 8 #75                 8 #80 Val Val Gln Arg Thr Pro Glu Glu Glu Arg As #n Leu Leu His Ala Asp                 885   #               890   #               895 Ser Asn Gly Tyr Thr Asn Leu Pro Asp Val Va #l Gln Pro Ser His Ser             900       #           905       #           910 Pro Thr Glu Asn Ser Lys Gly Gln Ser Pro Pr #o Ser Lys Asp Gly Ser         915           #       920           #       925 Gly Asp Tyr Gln Ser Arg Gly Leu Val Lys Al #a Pro Gly Lys Ser Ser     930               #   935               #   940 Phe Thr Met Phe Val Asp Leu Gly Ile Tyr Gl #n Pro Gly Gly Ser Gly 945                 9 #50                 9 #55                 9 #60 Asp Ser Ile Pro Ile Thr Ala Leu Val Gly Gl #y Glu Gly Thr Arg Leu                 965   #               970   #               975 Asp Gln Leu Gln Tyr Asp Val Arg Lys Gly Se #r Val Val Asn Val Asn             980       #           985       #           990 Pro Thr Asn Thr Arg Ala His Ser Glu Thr Pr #o Glu Ile Arg Lys Tyr         995           #      1000            #     1005 Lys Lys Arg Phe Asn Ser Glu Ile Leu Cys Al #a Ala Leu Trp Gly Val    1010               #  1015                # 1020 Asn Leu Leu Val Gly Thr Glu Asn Gly Leu Me #t Leu Leu Asp Arg Ser 1025               1030  #               1035   #              1040 Gly Gln Gly Lys Val Tyr Gly Leu Ile Gly Ar #g Arg Arg Phe Gln Gln                1045   #              1050    #             1055 Met Asp Val Leu Glu Gly Leu Asn Leu Leu Il #e Thr Ile Ser Gly Lys            1060       #          1065        #         1070 Arg Asn Lys Leu Arg Val Tyr Tyr Leu Ser Tr #p Leu Arg Asn Lys Ile        1075           #      1080            #     1085 Leu His Asn Asp Pro Glu Val Glu Lys Lys Gl #n Gly Trp Thr Thr Val    1090               #  1095                # 1100 Gly Asp Met Glu Gly Cys Gly His Tyr Arg Va #l Val Lys Tyr Glu Arg 1105               1110  #               1115   #              1120 Ile Lys Phe Leu Val Ile Ala Leu Lys Ser Se #r Val Glu Val Tyr Ala                1125   #              1130    #             1135 Trp Ala Pro Lys Pro Tyr His Lys Phe Met Al #a Phe Lys Ser Phe Ala            1140       #          1145        #         1150 Asp Leu Pro His Arg Pro Leu Leu Val Asp Le #u Thr Val Glu Glu Gly        1155           #      1160            #     1165 Gln Arg Leu Lys Val Ile Tyr Gly Ser Ser Al #a Gly Phe His Ala Val    1170               #  1175                # 1180 Asp Val Asp Ser Gly Asn Ser Tyr Asp Ile Ty #r Ile Pro Val His Ile 1185               1190  #               1195   #              1200 Gln Ser Gln Ile Thr Pro His Ala Ile Ile Ph #e Leu Pro Asn Thr Asp                1205   #              1210    #             1215 Gly Met Glu Met Leu Leu Cys Tyr Glu Asp Gl #u Gly Val Tyr Val Asn            1220       #          1225        #         1230 Thr Tyr Gly Arg Ile Ile Lys Asp Val Val Le #u Gln Trp Gly Glu Met        1235           #      1240            #     1245 Pro Thr Ser Val Ala Tyr Ile Cys Ser Asn Gl #n Ile Met Gly Trp Gly    1250               #  1255                # 1260 Glu Lys Ala Ile Glu Ile Arg Ser Val Glu Th #r Gly His Leu Asp Gly 1265               1270  #               1275   #              1280 Val Phe Met His Lys Arg Ala Gln Arg Leu Ly #s Phe Leu Cys Glu Arg                1285   #              1290    #             1295 Asn Asp Lys Val Phe Phe Ala Ser Val Arg Se #r Gly Gly Ser Ser Gln            1300       #          1305        #         1310 Val Tyr Phe Met Thr Leu Asn Arg Asn Arg Il #e Met Asn Trp        1315           #      1320            #     1325 <210> SEQ ID NO 16 <211> LENGTH: 349 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 16 Asn Val Asn Pro Leu Tyr Val Ser Pro Ala Cy #s Lys Lys Pro Leu Ile   1               5  #                 10  #                 15 His Met Tyr Glu Lys Glu Phe Thr Ser Glu Il #e Cys Cys Gly Ser Leu              20      #             25      #             30 Trp Gly Val Asn Leu Leu Leu Gly Thr Arg Se #r Asn Leu Tyr Leu Met          35          #         40          #         45 Asp Arg Ser Gly Lys Ala Asp Ile Thr Lys Le #u Ile Arg Arg Arg Pro      50              #     55              #     60 Phe Arg Gln Ile Gln Val Leu Glu Pro Leu As #n Leu Leu Ile Thr Ile  65                  # 70                  # 75                  # 80 Ser Gly His Lys Asn Arg Leu Arg Val Tyr Hi #s Leu Thr Trp Leu Arg                  85  #                 90  #                 95 Asn Lys Ile Leu Asn Asn Asp Pro Glu Ser Ly #s Arg Arg Gln Glu Glu             100       #           105       #           110 Met Leu Lys Thr Glu Glu Ala Cys Lys Ala Il #e Asp Lys Leu Thr Gly         115           #       120           #       125 Cys Glu His Phe Ser Val Leu Gln His Glu Gl #u Thr Thr Tyr Ile Ala     130               #   135               #   140 Ile Ala Leu Lys Ser Ser Ile His Leu Tyr Al #a Trp Ala Pro Lys Ser 145                 1 #50                 1 #55                 1 #60 Phe Asp Glu Ser Thr Ala Ile Lys Val Phe Pr #o Thr Leu Asp His Lys                 165   #               170   #               175 Pro Val Thr Val Asp Leu Ala Ile Gly Ser Gl #u Lys Arg Leu Lys Ile             180       #           185       #           190 Phe Phe Ser Ser Ala Asp Gly Tyr His Leu Il #e Asp Ala Glu Ser Glu         195           #       200           #       205 Val Met Ser Asp Val Thr Leu Pro Lys Asn Pr #o Leu Glu Ile Ile Ile     210               #   215               #   220 Pro Gln Asn Ile Ile Ile Leu Pro Asp Cys Le #u Gly Ile Gly Met Met 225                 2 #30                 2 #35                 2 #40 Leu Thr Phe Asn Ala Glu Ala Leu Ser Val Gl #u Ala Asn Glu Gln Leu                 245   #               250   #               255 Phe Lys Lys Ile Leu Glu Met Trp Lys Asp Il #e Pro Ser Ser Ile Ala             260       #           265       #           270 Phe Glu Cys Thr Gln Arg Thr Thr Gly Trp Gl #y Gln Lys Ala Ile Glu         275           #       280           #       285 Val Arg Ser Leu Gln Ser Arg Val Leu Glu Se #r Glu Leu Lys Arg Arg     290               #   295               #   300 Ser Ile Lys Lys Leu Arg Phe Leu Cys Thr Ar #g Gly Asp Lys Leu Phe 305                 3 #10                 3 #15                 3 #20 Phe Thr Ser Thr Leu Arg Asn His His Ser Ar #g Val Tyr Phe Met Thr                 325   #               330   #               335 Leu Gly Lys Leu Glu Glu Leu Gln Ser Asn Ty #r Asp Val             340       #           345 <210> SEQ ID NO 17 <211> LENGTH: 4023 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 17 ccgccatgaa ccccggcttc gatttgtccc gccggaaccc gcaggaggac tt #cgagctga     60 ttcagcgcat cggcagcggc acctacggcg acgtctacaa ggcacggaat gt #taacactg    120 gtgaattagc agcaattaaa gtaataaaat tggaaccagg agaagacttt gc #agttgtgc    180 agcaagaaat tattatgatg aaagactgta aacacccaaa tattgttgct ta #ttttggaa    240 gctatctcag gcgagataag ctttggattt gcatggagtt ttgtggaggt gg #ttctttac    300 aggatattta tcacgtaact ggacctctgt cagaactgca aattgcatat gt #tagcagag    360 aaacactgca gggattatat tatcttcaca gtaaaggaaa aatgcacaga ga #tataaagg    420 gagctaacat tctattaacg gataatggtc atgtgaaatt ggctgatttt gg #agtatctg    480 cacagataac agctacaatt gccaaacgga agtctttcat tggcacacca ta #ttggatgg    540 ctccagaagt tgcagctgtt gagaggaagg ggggttacaa tcaactctgt ga #tctctggg    600 cagtgggaat cactgccata gaacttgcag agcttcagcc tcctatgttt ga #cttacacc    660 caatgagagc attatttcta atgacaaaaa gcaattttca gcctcctaaa ct #aaaggata    720 aaatgaaatg gtcaaatagt tttcatcact ttgtgaaaat ggcacttacc aa #aaatccga    780 aaaaaagacc tactgctgaa aaattattac agcatccttt tgtaacacaa ca #tttgacac    840 ggtctttggc aatcgagctg ttggataaag taaataatcc agatcattcc ac #ttaccatg    900 atttcgatga tgatgatcct gagcctcttg ttgctgtacc acatagaatt ca #ctcaacaa    960 gtagaaacgt gagagaagaa aaaacacgct cagagataac ctttggccaa gt #gaaatttg   1020 atccaccctt aagaaaggag acagaaccac atcatgaact tcccgacagt ga #tggttttt   1080 tggacagttc agaagaaata tactacactg caagatctaa tctggatctg ca #actggaat   1140 atggacaagg acaccaaggt ggttactttt taggtgcaaa caagagtctt ct #caagtctg   1200 ttgaagaaga attgcatcag cgaggacacg tcgcacattt agaagatgat ga #aggagatg   1260 atgatgaatc taaacactca actctgaaag caaaaattcc acctcctttg cc #accaaagc   1320 ctaagtctat cttcatacca caggaaatgc attctactga ggatgaaaat ca #aggaacaa   1380 tcaagagatg tcccatgtca gggagcccag caaagccatc ccaagttcca cc #tagaccac   1440 cacctcccag attaccccca cacaaacctg ttgccttagg aaatggaatg ag #ctccttcc   1500 agttaaatgg tgaacgagat ggctcattat gtcaacaaca gaatgaacat ag #aggcacaa   1560 acctttcaag aaaagaaaag aaagatgtac caaagcctat tagtaatggt ct #tcctccaa   1620 cacctaaagt gcatatgggt gcatgttttt caaaagtttt taatgggtgt cc #cttgaaaa   1680 ttcactgtgc atcatcatgg ataaacccag atacaagaga tcagtacttg at #atttggtg   1740 ccgaagaagg gatttatacc ctcaatctta atgaacttca tgaaacatca at #ggaacagc   1800 tattccctcg aaggtgtaca tggttgtatg taatgaacaa ttgcttgcta tc #aatatctg   1860 gtaaagcttc tcagctttat tcccataatt taccagggct ttttgattat gc #aagacaaa   1920 tgcaaaagtt acctgttgct attccagcac acaaactccc tgacagaata ct #gccaagga   1980 aattttctgt atcagcaaaa atccctgaaa ccaaatggtg ccagaagtgt tg #tgttgtaa   2040 gaaatcctta cacgggccat aaatacctat gtggagcact tcagactagc at #tgttctat   2100 tagaatgggt tgaaccaatg cagaaattta tgttaattaa gcacatagat tt #tcctatac   2160 catgtccact tagaatgttt gaaatgctgg tagttcctga acaggagtac cc #tttagttt   2220 gtgttggtgt cagtagaggt agagacttca accaagtggt tcgatttgag ac #ggtcaatc   2280 caaattctac ctcttcatgg tttacagaat cagatacccc acagacaaat gt #tactcatg   2340 taacccaact ggagagagat accatccttg tatgcttgga ctgttgtata aa #aatagtaa   2400 atctccaagg aagattaaaa tctagcagga aattgtcatc agaactcacc tt #tgatttcc   2460 agattgaatc aatagtgtgc ctacaagaca gtgtgctagc tttctggaaa ca #tggaatgc   2520 aaggtagaag ttttagatct aatgaggtaa cacaagaaat ttcagatagc ac #aagaattt   2580 tcaggctgct tggatctgac agggtcgtgg ttttggaaag taggccaact ga #taacccca   2640 cagcaaatag caatttgtac atcctggcgg gtcatgaaaa cagttactga ga #attgttgt   2700 gctttgacag ttaactctag aaagaaagaa cactaccact gcaacattaa tg #gatgcttg   2760 aagctgtaca aaagctgcag taacctgtct tcagttactt tgtaatttat tg #tggcatga   2820 gataagatgg ggaaaatttt gttttaagtg gtatggatat atttagcata tt #gaaccaca   2880 caagtgctta attcattgtt atgtaatctt tgtacatata ggcagtattt tt #tctgtgaa   2940 acttcatatt gctgaagaca tacactaaga atttatgtag ataatgtact tt #tatgagat   3000 gtacaagtaa gtgtcttatc tgtacagatg taaatgttga tgaaaatgca at #tggggtta   3060 atattttaag aattctttag tatattcttg ggtgtggcta tattacaaaa tg #ggatgctg   3120 gcaatgaaac aatacattta acactattgt atttttatta tatgtaattt ag #taatatga   3180 atataaatct tgtaactttt aaaattgtaa tggaggctgt aatcatttta ta #atcttttt   3240 aattttaatg caagtacact ggtgtttata tttgcacaaa gtattgatat gt #gatgtatt   3300 aagtcacaaa agtaagctgt gacattgtct ataagcattt ggctccacaa at #gtatttgg   3360 attgttttct atgtgaagca aaccaattat aattaaccac atgttgtagt aa #ctggtctt   3420 tttatattta agcagaatcc tgtaagattg cttgtctttg cttaaaaaca at #acctttga   3480 acatttttga atcacagaat agcggtacca tgatagaata ctgcaattgt gg #tcagaatt   3540 acagtatgca caaagaatta attagcatta ttaaagagtc ctcactaaac at #ttcatatg   3600 atcacactga agaactgtaa cattccatag agtgaagtgg ttcaaatttc tc #ttggaatt   3660 tttacttttg ttggccttat tttatgatcc ttttcatatt tcttttgact ta #gagtatta   3720 atacatggcc aaaataattt agttactacc tcatacaaac aatataatgg tt #actacaca   3780 tcacaggaac ttagttttgg tttaagtcat ttttgattgc ttttttccaa tg #gaatatgt   3840 atataccagg ttttagcaaa atgcacactt ttggctcttt ttggtatatg tt #ctttatat   3900 tttaatgtga gtatatacac taagaacaaa ctaaattgtg atttatgatc tt #catttatt   3960 ttaatgataa tggttttaaa atatgttcct gattgtacat attgtaaaat aa #acatgttt   4020 ttt                   #                   #                   #           4023 <210> SEQ ID NO 18 <211> LENGTH: 894 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 18 Met Asn Pro Gly Phe Asp Leu Ser Arg Arg As #n Pro Gln Glu Asp Phe   1               5  #                 10  #                 15 Glu Leu Ile Gln Arg Ile Gly Ser Gly Thr Ty #r Gly Asp Val Tyr Lys              20      #             25      #             30 Ala Arg Asn Val Asn Thr Gly Glu Leu Ala Al #a Ile Lys Val Ile Lys          35          #         40          #         45 Leu Glu Pro Gly Glu Asp Phe Ala Val Val Gl #n Gln Glu Ile Ile Met      50              #     55              #     60 Met Lys Asp Cys Lys His Pro Asn Ile Val Al #a Tyr Phe Gly Ser Tyr  65                  # 70                  # 75                  # 80 Leu Arg Arg Asp Lys Leu Trp Ile Cys Met Gl #u Phe Cys Gly Gly Gly                  85  #                 90  #                 95 Ser Leu Gln Asp Ile Tyr His Val Thr Gly Pr #o Leu Ser Glu Leu Gln             100       #           105       #           110 Ile Ala Tyr Val Ser Arg Glu Thr Leu Gln Gl #y Leu Tyr Tyr Leu His         115           #       120           #       125 Ser Lys Gly Lys Met His Arg Asp Ile Lys Gl #y Ala Asn Ile Leu Leu     130               #   135               #   140 Thr Asp Asn Gly His Val Lys Leu Ala Asp Ph #e Gly Val Ser Ala Gln 145                 1 #50                 1 #55                 1 #60 Ile Thr Ala Thr Ile Ala Lys Arg Lys Ser Ph #e Ile Gly Thr Pro Tyr                 165   #               170   #               175 Trp Met Ala Pro Glu Val Ala Ala Val Glu Ar #g Lys Gly Gly Tyr Asn             180       #           185       #           190 Gln Leu Cys Asp Leu Trp Ala Val Gly Ile Th #r Ala Ile Glu Leu Ala         195           #       200           #       205 Glu Leu Gln Pro Pro Met Phe Asp Leu His Pr #o Met Arg Ala Leu Phe     210               #   215               #   220 Leu Met Thr Lys Ser Asn Phe Gln Pro Pro Ly #s Leu Lys Asp Lys Met 225                 2 #30                 2 #35                 2 #40 Lys Trp Ser Asn Ser Phe His His Phe Val Ly #s Met Ala Leu Thr Lys                 245   #               250   #               255 Asn Pro Lys Lys Arg Pro Thr Ala Glu Lys Le #u Leu Gln His Pro Phe             260       #           265       #           270 Val Thr Gln His Leu Thr Arg Ser Leu Ala Il #e Glu Leu Leu Asp Lys         275           #       280           #       285 Val Asn Asn Pro Asp His Ser Thr Tyr His As #p Phe Asp Asp Asp Asp     290               #   295               #   300 Pro Glu Pro Leu Val Ala Val Pro His Arg Il #e His Ser Thr Ser Arg 305                 3 #10                 3 #15                 3 #20 Asn Val Arg Glu Glu Lys Thr Arg Ser Glu Il #e Thr Phe Gly Gln Val                 325   #               330   #               335 Lys Phe Asp Pro Pro Leu Arg Lys Glu Thr Gl #u Pro His His Glu Leu             340       #           345       #           350 Pro Asp Ser Asp Gly Phe Leu Asp Ser Ser Gl #u Glu Ile Tyr Tyr Thr         355           #       360           #       365 Ala Arg Ser Asn Leu Asp Leu Gln Leu Glu Ty #r Gly Gln Gly His Gln     370               #   375               #   380 Gly Gly Tyr Phe Leu Gly Ala Asn Lys Ser Le #u Leu Lys Ser Val Glu 385                 3 #90                 3 #95                 4 #00 Glu Glu Leu His Gln Arg Gly His Val Ala Hi #s Leu Glu Asp Asp Glu                 405   #               410   #               415 Gly Asp Asp Asp Glu Ser Lys His Ser Thr Le #u Lys Ala Lys Ile Pro             420       #           425       #           430 Pro Pro Leu Pro Pro Lys Pro Lys Ser Ile Ph #e Ile Pro Gln Glu Met         435           #       440           #       445 His Ser Thr Glu Asp Glu Asn Gln Gly Thr Il #e Lys Arg Cys Pro Met     450               #   455               #   460 Ser Gly Ser Pro Ala Lys Pro Ser Gln Val Pr #o Pro Arg Pro Pro Pro 465                 4 #70                 4 #75                 4 #80 Pro Arg Leu Pro Pro His Lys Pro Val Ala Le #u Gly Asn Gly Met Ser                 485   #               490   #               495 Ser Phe Gln Leu Asn Gly Glu Arg Asp Gly Se #r Leu Cys Gln Gln Gln             500       #           505       #           510 Asn Glu His Arg Gly Thr Asn Leu Ser Arg Ly #s Glu Lys Lys Asp Val         515           #       520           #       525 Pro Lys Pro Ile Ser Asn Gly Leu Pro Pro Th #r Pro Lys Val His Met     530               #   535               #   540 Gly Ala Cys Phe Ser Lys Val Phe Asn Gly Cy #s Pro Leu Lys Ile His 545                 5 #50                 5 #55                 5 #60 Cys Ala Ser Ser Trp Ile Asn Pro Asp Thr Ar #g Asp Gln Tyr Leu Ile                 565   #               570   #               575 Phe Gly Ala Glu Glu Gly Ile Tyr Thr Leu As #n Leu Asn Glu Leu His             580       #           585       #           590 Glu Thr Ser Met Glu Gln Leu Phe Pro Arg Ar #g Cys Thr Trp Leu Tyr         595           #       600           #       605 Val Met Asn Asn Cys Leu Leu Ser Ile Ser Gl #y Lys Ala Ser Gln Leu     610               #   615               #   620 Tyr Ser His Asn Leu Pro Gly Leu Phe Asp Ty #r Ala Arg Gln Met Gln 625                 6 #30                 6 #35                 6 #40 Lys Leu Pro Val Ala Ile Pro Ala His Lys Le #u Pro Asp Arg Ile Leu                 645   #               650   #               655 Pro Arg Lys Phe Ser Val Ser Ala Lys Ile Pr #o Glu Thr Lys Trp Cys             660       #           665       #           670 Gln Lys Cys Cys Val Val Arg Asn Pro Tyr Th #r Gly His Lys Tyr Leu         675           #       680           #       685 Cys Gly Ala Leu Gln Thr Ser Ile Val Leu Le #u Glu Trp Val Glu Pro     690               #   695               #   700 Met Gln Lys Phe Met Leu Ile Lys His Ile As #p Phe Pro Ile Pro Cys 705                 7 #10                 7 #15                 7 #20 Pro Leu Arg Met Phe Glu Met Leu Val Val Pr #o Glu Gln Glu Tyr Pro                 725   #               730   #               735 Leu Val Cys Val Gly Val Ser Arg Gly Arg As #p Phe Asn Gln Val Val             740       #           745       #           750 Arg Phe Glu Thr Val Asn Pro Asn Ser Thr Se #r Ser Trp Phe Thr Glu         755           #       760           #       765 Ser Asp Thr Pro Gln Thr Asn Val Thr His Va #l Thr Gln Leu Glu Arg     770               #   775               #   780 Asp Thr Ile Leu Val Cys Leu Asp Cys Cys Il #e Lys Ile Val Asn Leu 785                 7 #90                 7 #95                 8 #00 Gln Gly Arg Leu Lys Ser Ser Arg Lys Leu Se #r Ser Glu Leu Thr Phe                 805   #               810   #               815 Asp Phe Gln Ile Glu Ser Ile Val Cys Leu Gl #n Asp Ser Val Leu Ala             820       #           825       #           830 Phe Trp Lys His Gly Met Gln Gly Arg Ser Ph #e Arg Ser Asn Glu Val         835           #       840           #       845 Thr Gln Glu Ile Ser Asp Ser Thr Arg Ile Ph #e Arg Leu Leu Gly Ser     850               #   855               #   860 Asp Arg Val Val Val Leu Glu Ser Arg Pro Th #r Asp Asn Pro Thr Ala 865                 8 #70                 8 #75                 8 #80 Asn Ser Asn Leu Tyr Ile Leu Ala Gly His Gl #u Asn Ser Tyr                 885   #               890 <210> SEQ ID NO 19 <211> LENGTH: 4196 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 19 gggagggtcc ttgtggcgcc gggcggcggg gtcctgcgtg gagagtggga cg #caacgccg     60 agaccgcgag cagaggctgc gcacagccgg atccggcact cagcgaccgg ac #ccaaggat    120 ccgccgggga acaagccaca ggagagcgac tcaggaacaa gtgtgggaga gg #aagcggcg    180 gcggcggcgc cgggcccggg ggtggtgaca gcaggtctga ggttgcatca ta #aatacaaa    240 ggactgaagt tataaaagag aaaagagaag tttgctgcta aaatgaatct ga #gcaatatg    300 gaatattttg tgccacacac aaaaaggtac tgaagattta ccccccaaaa aa #aattgtca    360 atgagaaata aagctaactg atatcaaaaa gcagagcctg ctctactggc ca #tcatgcgt    420 aaaggggtgc tgaaggaccc agagattgac gatctattct acaaagatga tc #ctgaggaa    480 ctttttattg gtttgcatga aattggacat ggaagttttg gagcagttta tt #ttgctaca    540 aatgctcaca ccaatgaggt ggtggcaatt aagaagatgt cctatagtgg ga #agcagacc    600 catgagaaat ggcaagatat tcttaaggaa gttaaatttt tacgacaatt ga #agcatcct    660 aatactattg agtacaaagg ctgttacttg aaagaacaca ctgcttggtt gg #tgatggaa    720 tattgcttag gctcagcctc tgatttatta gaagttcata aaaaaccact tc #aggaagtg    780 gagatcgctg ccattactca tggagccttg catggactag cctacctaca tt #ctcatgca    840 ttgattcata gggatattaa agcaggaaat attcttctaa cagagccagg tc #aggtaaaa    900 ctagctgatt ttggatctgc ttcaatggct tctcctgcca actccttcgt gg #gcacacct    960 tactggatgg ctccagaggt gatcttagct atggatgaag gacagtatga tg #ggaaagtt   1020 gatatttggt cacttggcat cacttgtatt gaattggcgg aacggaagcc gc #cccttttc   1080 aacatgaatg caatgagtgc cttatatcac attgcccaga atgactcccc aa #cgttacag   1140 tctaatgaat ggacagactc ctttaggaga tttgttgatt actgcttgca ga #aaatacct   1200 caggaaaggc caacatcagc agaactatta aggcatgact ttgttcgacg ag #accggcca   1260 ctacgtgtcc tcattgacct catacagagg acaaaagatg cagttcgtga gc #tagataac   1320 ctacagtacc gaaaaatgaa aaaaatactt ttccaagaga cacggaatgg ac #ccttgaat   1380 gagtcacagg aggatgagga agacagtgaa catggaacca gcctgaacag gg #aaatggac   1440 agcctgggca gcaaccattc cattccaagc atgtccgtga gcacaggcag cc #agagcagc   1500 agtgtgaaca gcatgcagga agtcatggac gagagcagtt ccgaacttgt ca #tgatgcac   1560 gatgacgaaa gcacaatcaa ttccagctcc tccgtcgtgc ataagaaaga tc #atgtattc   1620 acaagggatg aggcgggcca cggcgatccc aggcctgagc cgcggcctac cc #agtcagtt   1680 cagagccagg ccctccacta ccggaacaga gagcgctttg ccacgatcaa at #cagcatct   1740 ttggttacac gacagatcca tgagcatgag caggagaacg agttgcggga ac #agatgtca   1800 ggttataagc ggatgcggcg ccagcaccag aagcagctga tcgccctgga ga #acaagctg   1860 aaggctgaga tggacgagca ccgcctcaag ctacagaagg aggtggagac gc #atgccaac   1920 aactcgtcca tcgagctgga gaagctggcc aagaagcaag tggctatcat ag #aaaaggag   1980 gcaaaggtag ctgcagcaga tgagaagaag ttccagcaac agatcttggc cc #agcagaag   2040 aaagatttga caactttctt agaaagtcag aagaagcagt ataagatttg ta #aggaaaaa   2100 ataaaagagg aaatgaatga ggaccatagc acacccaaga aagagaagca ag #agcggatc   2160 tccaaacata aagagaactt gcagcacaca caggctgaag aggaagccca cc #ttctcact   2220 caacagagac tgtactacga caaaaattgt cgtttcttca agcggaaaat aa #tgatcaag   2280 cggcacgagg tggagcagca gaacattcgg gaggaactaa ataaaaagag ga #cccagaag   2340 gagatggagc atgccatgct aatccggcac gacgagtcca cccgagagct ag #agtacagg   2400 cagctgcaca cgttacagaa gctacgcatg gatctgatcc gtttacagca cc #agacggaa   2460 ctggaaaacc agctggagta caataagagg cgagaaagag aactgcacag aa #agcatgtc   2520 atgggacttc ggcaacagcc aaaaaactta aaggccatgg aaatgcaaat ta #aaaaacag   2580 tttcaggaca cttgcaaagt acagaccaaa cagtataaag cactcaagaa tc #accagttg   2640 gaagttactc caaagaatga gcacaaaaca atcttaaaga cactgaaaga tg #agcagaca   2700 agaaaacttg ccattttggc agagcagtat gaacagagta taaatgaaat ga #tggcctct   2760 caagcgttac ggctagatga ggctcaagaa gcagaatgcc aggccttgag gc #tacagctc   2820 cagcaggaaa tggagctgct caacgcctac cagagcaaaa tcaagatgca aa #cagaggca   2880 caacatgaac gtgagctcca gaagctagag cagagagtgt ctctgcgcag ag #cacacctt   2940 gagcagaaga ttgaagagga gctggctgcc cttcagaagg aacgcagcga ga #gaataaag   3000 aacctattgg aaaggcaaga gcgagagatt gaaacttttg acatggagag cc #tcagaatg   3060 ggatttggga atttggttac attagatttt cctaaggagg actacagatg ag #attaaatt   3120 ttttgccatt tacaaaaaaa aaaaaaaaaa agaaaacaga aaaaaattca ga #ccctgcaa   3180 aaccacattc cccattttaa cgggcgttgc tctcactctc tctctctctt ac #tcttactg   3240 acatcgtgtc ggactagtgc ctgtttattc ttactccatc aggggccccc tt #cctccccc   3300 cgtgtcaact ttcagtgctg gccaaaacct ggccgtctct tctattcaca gt #acacgtca   3360 cagtattgat gtgattcaaa atgtttcagt gaaaactttg gagacagttt ta #acaaaacc   3420 aataaaccaa caacaaaaaa agtggatgta tattgcttta agcaatcact ca #ttaccacc   3480 aatctgtgaa agtaaagcaa aaaataataa taataaatgc caagggggag ag #agacacaa   3540 tatccgcagc cttacacctt aactagctgc tgcattattt tattttattt ta #tttttttg   3600 gtatttattc atcaggaata aaaaaaacaa agttttatta aagattgaaa at #ttgataca   3660 ttttacagaa actaattgtg atgtacatat cagtggtgac atattattac tt #ttttgggg   3720 acggggggtg ggtggggtga agagatcttg tgatttttaa gaacctgctg gc #aagagttt   3780 aacttgtctt cagcatattc tgattgtatc ataatcattt tctgctgttg ca #gaggatgt   3840 gaatacactt aaggagctca cagaatccca gtagcacaaa ttgggctttg gc #aaatcgtg   3900 tattttgtgt atagaaggaa tttaaggaga ggtattactt attttcatat tg #tattttaa   3960 ctgtttctct gatcaaattt ttttacttcc tcctcctgtt cctccccacc tc #cctccttt   4020 tccagttcag tatttggagt tcaacactgt ctctcaatca gatcatcttg at #ctttttct   4080 ttatctccct tccccttcct aagtcccatt tcttggtcat aaatattgca tt #attcacac   4140 tttcaaactg tgtattttct tacaataaaa aatgatgaaa aaaaaaaaaa aa #aaaa       4196 <210> SEQ ID NO 20 <211> LENGTH: 3824 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 20 tattgaattg gcggaacgga agcctccttt atttaatatg aatgcaatga gt #gccttata     60 tcacatagcc caaaatgaat cccctacact acagtctaat gaatggtctg at #tattttcg    120 caactttgta gattcttgcc tccagaaaat ccctcaagat cgacctacat ca #gaggaact    180 tttaaagcac atatttgttc ttcgggagcg ccctgaaacc gtgttaatag at #ctcattca    240 gaggacaaag gatgcagtaa gagagctgga caatctgcag tatcgaaaga tg #aagaaact    300 ccttttccag gaggcacata atggaccagc agtagaagca caggaagaag aa #gaggaaca    360 agatcatggt gttggccgga caggaacagt taatagtgtt ggaagtaatc aa #tccattcc    420 cagcatgtcc atcagtgcca gcagccaaag cagtagtgtt aacagtcttc ca #gatgtctc    480 agatgacaag agtgagctag acatgatgga gggagaccac acagtgatgt ct #aacagttc    540 tgttatccat ttaaaaccag aggaagaaaa ttacagagaa gagggagatc ct #agaacaag    600 agcatcagat ccacaatctc caccccaagt atctcgtcac aaatcacact at #cgtaatcg    660 agaacacttt gctactatac ggacagcatc actggttacg aggcaaatgc aa #gaacatga    720 gcaggactct gagcttagag aacaaatgtc tggctataag cgaatgaggc ga #caacatca    780 aaagcaactg atgactctgg aaaacaagct aaaggctgag atggatgaac at #cgcctcag    840 attagacaaa gatcttgaaa ctcagcgtaa caattttgct gcagaaatgg ag #aaacttat    900 caagaaacac caggctgcca tggagaaaga ggctaaagtg atgtccaatg aa #gagaaaaa    960 atttcagcaa catattcagg cccaacagaa gaaagaactg aatagttttc tc #gagtccca   1020 gaaaagagag tataaacttc gaaaagagca gcttaaagag gagctaaatg aa #aaccagag   1080 tacccccaaa aaagaaaaac aggagtggct ttcaaagcag aaggagaata ta #cagcattt   1140 ccaagcagaa gaagaagcta accttcttcg acgtcaaaga caatacctag ag #ctggaatg   1200 ccgtcgcttc aagagaagaa tgttacttgg gcgtcataac ttagagcagg ac #cttgtcag   1260 ggaggagtta aacaaaagac agactcagaa ggacttagag catgccatgc ta #ctccgaca   1320 gcatgaatct atgcaagaac tggagttccg ccacctcaac acaattcaga ag #atgcgctg   1380 tgagttgatc agattacagc atcaaactga gctcactaac cagctggaat at #aataagcg   1440 aagagaacga gaactaagac gaaagcatgt catggaagtt cgacaacagc ct #aagagttt   1500 gaagtctaaa gaactccaaa taaaaaagca gtttcaggat acctgcaaaa tc #caaaccag   1560 acagtacaaa gcattaagaa atcacctgct ggagactaca ccaaagagtg ag #cacaaagc   1620 tgttctgaaa cggctcaagg aggaacagac ccggaaatta gctatcttgg ct #gagcagta   1680 tgatcacagc attaatgaaa tgctctccac acaagccctg cgtttggatg aa #gcacagga   1740 agcagagtgc caggttttga agatgcagct gcagcaggaa ctggagctgt tg #aatgcgta   1800 tcagagcaaa atcaagatgc aagctgaggc acaacatgat cgagagcttc gc #gagcttga   1860 acagagggtc tccctccgga gggcactctt agaacaaaag attgaagaag ag #atgttggc   1920 tttgcagaat gagcgcacag aacgaatacg aagcctgttg gaacgtcaag cc #agagagat   1980 tgaagctttt gactctgaaa gcatgagact aggttttagt aatatggtcc tt #tctaatct   2040 ctcccctgag gcattcagcc acagctaccc gggagcttct ggttggtcac ac #aaccctac   2100 tgggggtcca ggacctcact ggggtcatcc catgggtggc ccaccacaag ct #tggggcca   2160 tccaatgcaa ggtggacccc agccatgggg tcacccttca gggccaatgc aa #ggggtacc   2220 tcgaggtagc agtatgggag tccgcaatag cccccaggct ctgaggcgga ca #gcttctgg   2280 gggacggacg gagcagggca tgagcagaag cacgagtgtc acttcacaaa ta #tccaatgg   2340 gtcacacatg tcttatacat aacttaataa ttgagagtgg caattccgct gg #agctgtct   2400 gccaaaagaa actgcctaca gacatcatca cagcagcctc ctcacttggg ta #ctacagtg   2460 tggaagctga gtgcatatgg tatattttat tcatttttgt aaagcgttct gt #tttgtgtt   2520 tactaattgg gatgtcatag tacttggctg ccgggtttgt ttgtttttgg gg #aaattttg   2580 aaaagtggag ttgatattaa aaataaatgt gtatgtgtgt acatatatat ac #acacacat   2640 acacatatat tatgcatgtg gtgaaaagaa ttggctagat aggggatttt tc #tgaacact   2700 gcaaaaatag aacgtagcaa aatggcttca gttatcactt ttgggtgtct gt #atcctaag   2760 aagtttctga aaagatctaa agccttttta tcccatatcc caaattctta tg #agccactc   2820 acagcaggca gcatatgttg aaataagtta ttactggtac acacctgcat tg #cctcacca   2880 gtgtatttat ttgttattaa attgatctga cttctcagcc tcatttggac ta #aaaaaaga   2940 aagcagaaat ccatgaacac attgcttctc ggccttttgg ctaagatcaa gt #gtagaaat   3000 ccatgaacac taaaggactt cattgatttt ttcagagagt agaaaacaac tt #agtttttc   3060 ttttttcctg aatgcgtcat aggcttgtga gtgatttttg tccattcaat tg #tgccttct   3120 ttgtattatg ataagatggg ggtacttaag gagatcacaa gttgtgtgag ga #ttgcatta   3180 acaaacctat gagccttcaa tggggaagac cagaagggtg agaggggccc tg #aaagttca   3240 tatggtgggt atgtcccgca gcagagtgag gagatgaagc ttacgtgtcc tg #acgttttg   3300 ttgcttatac tgtgatatct catcctagct aagctctata atgcccaaga cc #ccaaacag   3360 tacttttact ttgtttgtac aaaaacaaag acatatagcc aatacaaatc aa #atgccgga   3420 ggtgtttgat gccatatttg caaattgcca tctattgaaa ttctcgtcac ac #tacataga   3480 cataattgtt atctcctttt ggcttatgtg attttctgtt tacaagtaga at #agccaatt   3540 atttaaatgt ttagttgcca cagtgaacca ggagtcactg agccaatgac tt #taccagct   3600 gctgactaat cttcatcacc actgtagatt ttgctgcatg tgcaggtcct ct #atttttaa   3660 ttgctgtttt cgttgctgca gtactttaca aacttctagt tcgttgagac tt #agtgacca   3720 tttggcatca agttaacatc acacaatagg aaacaccact tccacaagtc tc #aagcctca   3780 gtgctaaagt actactgaaa aggaactagg aagtttggcc aatt    #                 382 #4 <210> SEQ ID NO 21 <211> LENGTH: 2249 <212> TYPE: DNA <213> ORGANISM: Murine sp. <400> SEQUENCE: 21 gcaggatgcc atcaactaac agagcaggca gtctaaagga ccctgaaatt gc #agagctct     60 tcttcaaaga agatccggaa aagctcttca cagatctcag agaaatcggc ca #tgggagct    120 ttggagcagt atattttgca cgagatgtgc gtactaatga agtggtggcc at #caagaaaa    180 tgtcttatag tggaaagcag tctactgaga aatggcagga tattattaag ga #agtcaagt    240 ttctacaaag aataaaacat cccaacagta tagaatacaa aggctgctat tt #acgtgaac    300 acacagcatg gcttgtaatg gaatattgtt taggatctgc ttcagattta tt #agaagttc    360 ataaaaagcc attacaagaa gtggaaatag cagcaattac acatggtgct ct #ccagggac    420 tagcttattt acattctcat accatgatcc atagagatat caaagcagga aa #tatccttc    480 tgacagaacc aggccaagtg aaacttgctg actttggatc tgcttccatg gc #ttcccctg    540 ccaattcttt tgtgggaaca ccatattgga tggccccaga agtaatttta gc #catggatg    600 aaggacagta tgatggcaaa gttgatgtat ggtctcttgg aataacgtgt at #tgaattag    660 ccgagaggaa gcctccttta tttaatatga atgcaatgag tgccttatat ca #catagccc    720 aaaatgaatc ccctacacta caatctaata tgaatgattc ttgcctccag aa #aatccctc    780 aagatcgccc tacatcagag gaacttttaa agcacatgtt tgttcttcga ga #gcgccctg    840 aaacagtgtt aatagatctt attcaaagga caaaggatgc agtaagagag ct #ggacaatc    900 tgcagtatcg aaagatgaag aaactccttt tccaggaggc acataatggg cc #agcggtag    960 aagcacagga agaagaggag gagcaagatc atggtgttgg ccgaacagga ac #agtgaata   1020 gtgttggaag caatcagtct atccctagta tgtctatcag tgccagcagt ca #aagcagca   1080 gtgttaatag tcttccagat gcatcagatg acaagagtga gctagacatg at #ggagggag   1140 accatacagt gatgtctaac agttctgtca tccacttaaa acctgaggag ga #aaattacc   1200 aggaagaagg agatcctaga acaagagcat cagacccaca gtctccccct ca #ggtgtctc   1260 gtcacaagtc acattatcgt aatagagaac actttgcaac catacgaaca gc #atcactgg   1320 ttacaagaca gatgcaagaa catgagcagg actctgaact tagagaacag at #gtctggtt   1380 ataagcggat gaggcgacag catcaaaagc agctgatgac gctggaaaat aa #actgaagg   1440 cagagatgga cgaacatcgg ctcagattag acaaagatct tgaaactcag cg #taacaatt   1500 tcgctgcaga aatggagaaa cttattaaga aacaccaagc tgctatggaa aa #agaggcta   1560 aagtgatggc caatgaggag aaaaaattcc agcaacacat tcaggctcaa ca #gaaaaaag   1620 aactgaatag ctttttggag tctcaaaaaa gagaatataa acttcgcaaa ga #gcagctta   1680 aggaggagct gaatgaaaac cagagcacac ctaaaaaaga aaagcaggaa tg #gctttcaa   1740 agcagaagga gaatatacag cattttcagg cagaagaaga agctaatctt ct #tcgacgtc   1800 aaaggcagta tctagagcta gaatgtcgtc gcttcaaaag aagaatgtta ct #tgggcgac   1860 ataacttgga acaggacctt gtcagggagg agttaaacaa aaggcagact ca #aaaggact   1920 tggaacatgc aatgctattg cgacagcatg aatcaatgca agaactggag tt #tcgccatc   1980 tcaacactat tcagaagatg cgctgtgagt tgatcagact gcagcatcaa ac #tgagctca   2040 ctaaccagct agagtacaat aagagaaggg aacgggaact gaggcgaaaa ca #tgtcatgg   2100 aagttcgaca acaacctaag agtctgaagt ctaaagaact ccaaataaaa aa #gcagtttc   2160 aggatacctg caaaattcaa accagacagt acaaagcatt aaggaatcac ct #actggaga   2220 ctacaccaaa gaatgagcac aaagcaatc          #                   #          2249 <210> SEQ ID NO 22 <211> LENGTH: 898 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 22 Met Arg Lys Gly Val Leu Lys Asp Pro Glu Il #e Asp Asp Leu Phe Tyr   1               5  #                 10  #                 15 Lys Asp Asp Pro Glu Glu Leu Phe Ile Gly Le #u His Glu Ile Gly His              20      #             25      #             30 Gly Ser Phe Gly Ala Val Tyr Phe Ala Thr As #n Ala His Thr Asn Glu          35          #         40          #         45 Val Val Ala Ile Lys Lys Met Ser Tyr Ser Gl #y Lys Gln Thr His Glu      50              #     55              #     60 Lys Trp Gln Asp Ile Leu Lys Glu Val Lys Ph #e Leu Arg Gln Leu Lys  65                  # 70                  # 75                  # 80 His Pro Asn Thr Ile Glu Tyr Lys Gly Cys Ty #r Leu Lys Glu His Thr                  85  #                 90  #                 95 Ala Trp Leu Val Met Glu Tyr Cys Leu Gly Se #r Ala Ser Asp Leu Leu             100       #           105       #           110 Glu Val His Lys Lys Pro Leu Gln Glu Val Gl #u Ile Ala Ala Ile Thr         115           #       120           #       125 His Gly Ala Leu His Gly Leu Ala Tyr Leu Hi #s Ser His Ala Leu Ile     130               #   135               #   140 His Arg Asp Ile Lys Ala Gly Asn Ile Leu Le #u Thr Glu Pro Gly Gln 145                 1 #50                 1 #55                 1 #60 Val Lys Leu Ala Asp Phe Gly Ser Ala Ser Me #t Ala Ser Pro Ala Asn                 165   #               170   #               175 Ser Phe Val Gly Thr Pro Tyr Trp Met Ala Pr #o Glu Val Ile Leu Ala             180       #           185       #           190 Met Asp Glu Gly Gln Tyr Asp Gly Lys Val As #p Ile Trp Ser Leu Gly         195           #       200           #       205 Ile Thr Cys Ile Glu Leu Ala Glu Arg Lys Pr #o Pro Leu Phe Asn Met     210               #   215               #   220 Asn Ala Met Ser Ala Leu Tyr His Ile Ala Gl #n Asn Asp Ser Pro Thr 225                 2 #30                 2 #35                 2 #40 Leu Gln Ser Asn Glu Trp Thr Asp Ser Phe Ar #g Arg Phe Val Asp Tyr                 245   #               250   #               255 Cys Leu Gln Lys Ile Pro Gln Glu Arg Pro Th #r Ser Ala Glu Leu Leu             260       #           265       #           270 Arg His Asp Phe Val Arg Arg Asp Arg Pro Le #u Arg Val Leu Ile Asp         275           #       280           #       285 Leu Ile Gln Arg Thr Lys Asp Ala Val Arg Gl #u Leu Asp Asn Leu Gln     290               #   295               #   300 Tyr Arg Lys Met Lys Lys Ile Leu Phe Gln Gl #u Thr Arg Asn Gly Pro 305                 3 #10                 3 #15                 3 #20 Leu Asn Glu Ser Gln Glu Asp Glu Glu Asp Se #r Glu His Gly Thr Ser                 325   #               330   #               335 Leu Asn Arg Glu Met Asp Ser Leu Gly Ser As #n His Ser Ile Pro Ser             340       #           345       #           350 Met Ser Val Ser Thr Gly Ser Gln Ser Ser Se #r Val Asn Ser Met Gln         355           #       360           #       365 Glu Val Met Asp Glu Ser Ser Ser Glu Leu Va #l Met Met His Asp Asp     370               #   375               #   380 Glu Ser Thr Ile Asn Ser Ser Ser Ser Val Va #l His Lys Lys Asp His 385                 3 #90                 3 #95                 4 #00 Val Phe Thr Arg Asp Glu Ala Gly His Gly As #p Pro Arg Pro Glu Pro                 405   #               410   #               415 Arg Pro Thr Gln Ser Val Gln Ser Gln Ala Le #u His Tyr Arg Asn Arg             420       #           425       #           430 Glu Arg Phe Ala Thr Ile Lys Ser Ala Ser Le #u Val Thr Arg Gln Ile         435           #       440           #       445 His Glu His Glu Gln Glu Asn Glu Leu Arg Gl #u Gln Met Ser Gly Tyr     450               #   455               #   460 Lys Arg Met Arg Arg Gln His Gln Lys Gln Le #u Ile Ala Leu Glu Asn 465                 4 #70                 4 #75                 4 #80 Lys Leu Lys Ala Glu Met Asp Glu His Arg Le #u Lys Leu Gln Lys Glu                 485   #               490   #               495 Val Glu Thr His Ala Asn Asn Ser Ser Ile Gl #u Leu Glu Lys Leu Ala             500       #           505       #           510 Lys Lys Gln Val Ala Ile Ile Glu Lys Glu Al #a Lys Val Ala Ala Ala         515           #       520           #       525 Asp Glu Lys Lys Phe Gln Gln Gln Ile Leu Al #a Gln Gln Lys Lys Asp     530               #   535               #   540 Leu Thr Thr Phe Leu Glu Ser Gln Lys Lys Gl #n Tyr Lys Ile Cys Lys 545                 5 #50                 5 #55                 5 #60 Glu Lys Ile Lys Glu Glu Met Asn Glu Asp Hi #s Ser Thr Pro Lys Lys                 565   #               570   #               575 Glu Lys Gln Glu Arg Ile Ser Lys His Lys Gl #u Asn Leu Gln His Thr             580       #           585       #           590 Gln Ala Glu Glu Glu Ala His Leu Leu Thr Gl #n Gln Arg Leu Tyr Tyr         595           #       600           #       605 Asp Lys Asn Cys Arg Phe Phe Lys Arg Lys Il #e Met Ile Lys Arg His     610               #   615               #   620 Glu Val Glu Gln Gln Asn Ile Arg Glu Glu Le #u Asn Lys Lys Arg Thr 625                 6 #30                 6 #35                 6 #40 Gln Lys Glu Met Glu His Ala Met Leu Ile Ar #g His Asp Glu Ser Thr                 645   #               650   #               655 Arg Glu Leu Glu Tyr Arg Gln Leu His Thr Le #u Gln Lys Leu Arg Met             660       #           665       #           670 Asp Leu Ile Arg Leu Gln His Gln Thr Glu Le #u Glu Asn Gln Leu Glu         675           #       680           #       685 Tyr Asn Lys Arg Arg Glu Arg Glu Leu His Ar #g Lys His Val Met Gly     690               #   695               #   700 Leu Arg Gln Gln Pro Lys Asn Leu Lys Ala Me #t Glu Met Gln Ile Lys 705                 7 #10                 7 #15                 7 #20 Lys Gln Phe Gln Asp Thr Cys Lys Val Gln Th #r Lys Gln Tyr Lys Ala                 725   #               730   #               735 Leu Lys Asn His Gln Leu Glu Val Thr Pro Ly #s Asn Glu His Lys Thr             740       #           745       #           750 Ile Leu Lys Thr Leu Lys Asp Glu Gln Thr Ar #g Lys Leu Ala Ile Leu         755           #       760           #       765 Ala Glu Gln Tyr Glu Gln Ser Ile Asn Glu Me #t Met Ala Ser Gln Ala     770               #   775               #   780 Leu Arg Leu Asp Glu Ala Gln Glu Ala Glu Cy #s Gln Ala Leu Arg Leu 785                 7 #90                 7 #95                 8 #00 Gln Leu Gln Gln Glu Met Glu Leu Leu Asn Al #a Tyr Gln Ser Lys Ile                 805   #               810   #               815 Lys Met Gln Thr Glu Ala Gln His Glu Arg Gl #u Leu Gln Lys Leu Glu             820       #           825       #           830 Gln Arg Val Ser Leu Arg Arg Ala His Leu Gl #u Gln Lys Ile Glu Glu         835           #       840           #       845 Glu Leu Ala Ala Leu Gln Lys Glu Arg Ser Gl #u Arg Ile Lys Asn Leu     850               #   855               #   860 Leu Glu Arg Gln Glu Arg Glu Ile Glu Thr Ph #e Asp Met Glu Ser Leu 865                 8 #70                 8 #75                 8 #80 Arg Met Gly Phe Gly Asn Leu Val Thr Leu As #p Phe Pro Lys Glu Asp                 885   #               890   #               895 Tyr Arg <210> SEQ ID NO 23 <211> LENGTH: 786 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 23 Ile Glu Leu Ala Glu Arg Lys Pro Pro Leu Ph #e Asn Met Asn Ala Met   1               5  #                 10  #                 15 Ser Ala Leu Tyr His Ile Ala Gln Asn Glu Se #r Pro Thr Leu Gln Ser              20      #             25      #             30 Asn Glu Trp Ser Asp Tyr Phe Arg Asn Phe Va #l Asp Ser Cys Leu Gln          35          #         40          #         45 Lys Ile Pro Gln Asp Arg Pro Thr Ser Glu Gl #u Leu Leu Lys His Ile      50              #     55              #     60 Phe Val Leu Arg Glu Arg Pro Glu Thr Val Le #u Ile Asp Leu Ile Gln  65                  # 70                  # 75                  # 80 Arg Thr Lys Asp Ala Val Arg Glu Leu Asp As #n Leu Gln Tyr Arg Lys                  85  #                 90  #                 95 Met Lys Lys Leu Leu Phe Gln Glu Ala His As #n Gly Pro Ala Val Glu             100       #           105       #           110 Ala Gln Glu Glu Glu Glu Glu Gln Asp His Gl #y Val Gly Arg Thr Gly         115           #       120           #       125 Thr Val Asn Ser Val Gly Ser Asn Gln Ser Il #e Pro Ser Met Ser Ile     130               #   135               #   140 Ser Ala Ser Ser Gln Ser Ser Ser Val Asn Se #r Leu Pro Asp Val Ser 145                 1 #50                 1 #55                 1 #60 Asp Asp Lys Ser Glu Leu Asp Met Met Glu Gl #y Asp His Thr Val Met                 165   #               170   #               175 Ser Asn Ser Ser Val Ile His Leu Lys Pro Gl #u Glu Glu Asn Tyr Arg             180       #           185       #           190 Glu Glu Gly Asp Pro Arg Thr Arg Ala Ser As #p Pro Gln Ser Pro Pro         195           #       200           #       205 Gln Val Ser Arg His Lys Ser His Tyr Arg As #n Arg Glu His Phe Ala     210               #   215               #   220 Thr Ile Arg Thr Ala Ser Leu Val Thr Arg Gl #n Met Gln Glu His Glu 225                 2 #30                 2 #35                 2 #40 Gln Asp Ser Glu Leu Arg Glu Gln Met Ser Gl #y Tyr Lys Arg Met Arg                 245   #               250   #               255 Arg Gln His Gln Lys Gln Leu Met Thr Leu Gl #u Asn Lys Leu Lys Ala             260       #           265       #           270 Glu Met Asp Glu His Arg Leu Arg Leu Asp Ly #s Asp Leu Glu Thr Gln         275           #       280           #       285 Arg Asn Asn Phe Ala Ala Glu Met Glu Lys Le #u Ile Lys Lys His Gln     290               #   295               #   300 Ala Ala Met Glu Lys Glu Ala Lys Val Met Se #r Asn Glu Glu Lys Lys 305                 3 #10                 3 #15                 3 #20 Phe Gln Gln His Ile Gln Ala Gln Gln Lys Ly #s Glu Leu Asn Ser Phe                 325   #               330   #               335 Leu Glu Ser Gln Lys Arg Glu Tyr Lys Leu Ar #g Lys Glu Gln Leu Lys             340       #           345       #           350 Glu Glu Leu Asn Glu Asn Gln Ser Thr Pro Ly #s Lys Glu Lys Gln Glu         355           #       360           #       365 Trp Leu Ser Lys Gln Lys Glu Asn Ile Gln Hi #s Phe Gln Ala Glu Glu     370               #   375               #   380 Glu Ala Asn Leu Leu Arg Arg Gln Arg Gln Ty #r Leu Glu Leu Glu Cys 385                 3 #90                 3 #95                 4 #00 Arg Arg Phe Lys Arg Arg Met Leu Leu Gly Ar #g His Asn Leu Glu Gln                 405   #               410   #               415 Asp Leu Val Arg Glu Glu Leu Asn Lys Arg Gl #n Thr Gln Lys Asp Leu             420       #           425       #           430 Glu His Ala Met Leu Leu Arg Gln His Glu Se #r Met Gln Glu Leu Glu         435           #       440           #       445 Phe Arg His Leu Asn Thr Ile Gln Lys Met Ar #g Cys Glu Leu Ile Arg     450               #   455               #   460 Leu Gln His Gln Thr Glu Leu Thr Asn Gln Le #u Glu Tyr Asn Lys Arg 465                 4 #70                 4 #75                 4 #80 Arg Glu Arg Glu Leu Arg Arg Lys His Val Me #t Glu Val Arg Gln Gln                 485   #               490   #               495 Pro Lys Ser Leu Lys Ser Lys Glu Leu Gln Il #e Lys Lys Gln Phe Gln             500       #           505       #           510 Asp Thr Cys Lys Ile Gln Thr Arg Gln Tyr Ly #s Ala Leu Arg Asn His         515           #       520           #       525 Leu Leu Glu Thr Thr Pro Lys Ser Glu His Ly #s Ala Val Leu Lys Arg     530               #   535               #   540 Leu Lys Glu Glu Gln Thr Arg Lys Leu Ala Il #e Leu Ala Glu Gln Tyr 545                 5 #50                 5 #55                 5 #60 Asp His Ser Ile Asn Glu Met Leu Ser Thr Gl #n Ala Leu Arg Leu Asp                 565   #               570   #               575 Glu Ala Gln Glu Ala Glu Cys Gln Val Leu Ly #s Met Gln Leu Gln Gln             580       #           585       #           590 Glu Leu Glu Leu Leu Asn Ala Tyr Gln Ser Ly #s Ile Lys Met Gln Ala         595           #       600           #       605 Glu Ala Gln His Asp Arg Glu Leu Arg Glu Le #u Glu Gln Arg Val Ser     610               #   615               #   620 Leu Arg Arg Ala Leu Leu Glu Gln Lys Ile Gl #u Glu Glu Met Leu Ala 625                 6 #30                 6 #35                 6 #40 Leu Gln Asn Glu Arg Thr Glu Arg Ile Arg Se #r Leu Leu Glu Arg Gln                 645   #               650   #               655 Ala Arg Glu Ile Glu Ala Phe Asp Ser Glu Se #r Met Arg Leu Gly Phe             660       #           665       #           670 Ser Asn Met Val Leu Ser Asn Leu Ser Pro Gl #u Ala Phe Ser His Ser         675           #       680           #       685 Tyr Pro Gly Ala Ser Gly Trp Ser His Asn Pr #o Thr Gly Gly Pro Gly     690               #   695               #   700 Pro His Trp Gly His Pro Met Gly Gly Pro Pr #o Gln Ala Trp Gly His 705                 7 #10                 7 #15                 7 #20 Pro Met Gln Gly Gly Pro Gln Pro Trp Gly Hi #s Pro Ser Gly Pro Met                 725   #               730   #               735 Gln Gly Val Pro Arg Gly Ser Ser Met Gly Va #l Arg Asn Ser Pro Gln             740       #           745       #           750 Ala Leu Arg Arg Thr Ala Ser Gly Gly Arg Th #r Glu Gln Gly Met Ser         755           #       760           #       765 Arg Ser Thr Ser Val Thr Ser Gln Ile Ser As #n Gly Ser His Met Ser     770               #   775               #   780 Tyr Thr 785 <210> SEQ ID NO 24 <211> LENGTH: 748 <212> TYPE: PRT <213> ORGANISM: Murine sp. <400> SEQUENCE: 24 Met Pro Ser Thr Asn Arg Ala Gly Ser Leu Ly #s Asp Pro Glu Ile Ala   1               5  #                 10  #                 15 Glu Leu Phe Phe Lys Glu Asp Pro Glu Lys Le #u Phe Thr Asp Leu Arg              20      #             25      #             30 Glu Ile Gly His Gly Ser Phe Gly Ala Val Ty #r Phe Ala Arg Asp Val          35          #         40          #         45 Arg Thr Asn Glu Val Val Ala Ile Lys Lys Me #t Ser Tyr Ser Gly Lys      50              #     55              #     60 Gln Ser Thr Glu Lys Trp Gln Asp Ile Ile Ly #s Glu Val Lys Phe Leu  65                  # 70                  # 75                  # 80 Gln Arg Ile Lys His Pro Asn Ser Ile Glu Ty #r Lys Gly Cys Tyr Leu                  85  #                 90  #                 95 Arg Glu His Thr Ala Trp Leu Val Met Glu Ty #r Cys Leu Gly Ser Ala             100       #           105       #           110 Ser Asp Leu Leu Glu Val His Lys Lys Pro Le #u Gln Glu Val Glu Ile         115           #       120           #       125 Ala Ala Ile Thr His Gly Ala Leu Gln Gly Le #u Ala Tyr Leu His Ser     130               #   135               #   140 His Thr Met Ile His Arg Asp Ile Lys Ala Gl #y Asn Ile Leu Leu Thr 145                 1 #50                 1 #55                 1 #60 Glu Pro Gly Gln Val Lys Leu Ala Asp Phe Gl #y Ser Ala Ser Met Ala                 165   #               170   #               175 Ser Pro Ala Asn Ser Phe Val Gly Thr Pro Ty #r Trp Met Ala Pro Glu             180       #           185       #           190 Val Ile Leu Ala Met Asp Glu Gly Gln Tyr As #p Gly Lys Val Asp Val         195           #       200           #       205 Trp Ser Leu Gly Ile Thr Cys Ile Glu Leu Al #a Glu Arg Lys Pro Pro     210               #   215               #   220 Leu Phe Asn Met Asn Ala Met Ser Ala Leu Ty #r His Ile Ala Gln Asn 225                 2 #30                 2 #35                 2 #40 Glu Ser Pro Thr Leu Gln Ser Asn Met Asn As #p Ser Cys Leu Gln Lys                 245   #               250   #               255 Ile Pro Gln Asp Arg Pro Thr Ser Glu Glu Le #u Leu Lys His Met Phe             260       #           265       #           270 Val Leu Arg Glu Arg Pro Glu Thr Val Leu Il #e Asp Leu Ile Gln Arg         275           #       280           #       285 Thr Lys Asp Ala Val Arg Glu Leu Asp Asn Le #u Gln Tyr Arg Lys Met     290               #   295               #   300 Lys Lys Leu Leu Phe Gln Glu Ala His Asn Gl #y Pro Ala Val Glu Ala 305                 3 #10                 3 #15                 3 #20 Gln Glu Glu Glu Glu Glu Gln Asp His Gly Va #l Gly Arg Thr Gly Thr                 325   #               330   #               335 Val Asn Ser Val Gly Ser Asn Gln Ser Ile Pr #o Ser Met Ser Ile Ser             340       #           345       #           350 Ala Ser Ser Gln Ser Ser Ser Val Asn Ser Le #u Pro Asp Ala Ser Asp         355           #       360           #       365 Asp Lys Ser Glu Leu Asp Met Met Glu Gly As #p His Thr Val Met Ser     370               #   375               #   380 Asn Ser Ser Val Ile His Leu Lys Pro Glu Gl #u Glu Asn Tyr Gln Glu 385                 3 #90                 3 #95                 4 #00 Glu Gly Asp Pro Arg Thr Arg Ala Ser Asp Pr #o Gln Ser Pro Pro Gln                 405   #               410   #               415 Val Ser Arg His Lys Ser His Tyr Arg Asn Ar #g Glu His Phe Ala Thr             420       #           425       #           430 Ile Arg Thr Ala Ser Leu Val Thr Arg Gln Me #t Gln Glu His Glu Gln         435           #       440           #       445 Asp Ser Glu Leu Arg Glu Gln Met Ser Gly Ty #r Lys Arg Met Arg Arg     450               #   455               #   460 Gln His Gln Lys Gln Leu Met Thr Leu Glu As #n Lys Leu Lys Ala Glu 465                 4 #70                 4 #75                 4 #80 Met Asp Glu His Arg Leu Arg Leu Asp Lys As #p Leu Glu Thr Gln Arg                 485   #               490   #               495 Asn Asn Phe Ala Ala Glu Met Glu Lys Leu Il #e Lys Lys His Gln Ala             500       #           505       #           510 Ala Met Glu Lys Glu Ala Lys Val Met Ala As #n Glu Glu Lys Lys Phe         515           #       520           #       525 Gln Gln His Ile Gln Ala Gln Gln Lys Lys Gl #u Leu Asn Ser Phe Leu     530               #   535               #   540 Glu Ser Gln Lys Arg Glu Tyr Lys Leu Arg Ly #s Glu Gln Leu Lys Glu 545                 5 #50                 5 #55                 5 #60 Glu Leu Asn Glu Asn Gln Ser Thr Pro Lys Ly #s Glu Lys Gln Glu Trp                 565   #               570   #               575 Leu Ser Lys Gln Lys Glu Asn Ile Gln His Ph #e Gln Ala Glu Glu Glu             580       #           585       #           590 Ala Asn Leu Leu Arg Arg Gln Arg Gln Tyr Le #u Glu Leu Glu Cys Arg         595           #       600           #       605 Arg Phe Lys Arg Arg Met Leu Leu Gly Arg Hi #s Asn Leu Glu Gln Asp     610               #   615               #   620 Leu Val Arg Glu Glu Leu Asn Lys Arg Gln Th #r Gln Lys Asp Leu Glu 625                 6 #30                 6 #35                 6 #40 His Ala Met Leu Leu Arg Gln His Glu Ser Me #t Gln Glu Leu Glu Phe                 645   #               650   #               655 Arg His Leu Asn Thr Ile Gln Lys Met Arg Cy #s Glu Leu Ile Arg Leu             660       #           665       #           670 Gln His Gln Thr Glu Leu Thr Asn Gln Leu Gl #u Tyr Asn Lys Arg Arg         675           #       680           #       685 Glu Arg Glu Leu Arg Arg Lys His Val Met Gl #u Val Arg Gln Gln Pro     690               #   695               #   700 Lys Ser Leu Lys Ser Lys Glu Leu Gln Ile Ly #s Lys Gln Phe Gln Asp 705                 7 #10                 7 #15                 7 #20 Thr Cys Lys Ile Gln Thr Arg Gln Tyr Lys Al #a Leu Arg Asn His Leu                 725   #               730   #               735 Leu Glu Thr Thr Pro Lys Asn Glu His Lys Al #a Ile             740       #           745 <210> SEQ ID NO 25 <211> LENGTH: 2795 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 25 cgaagccaca gcccgagccc gagcccgagc ccgagccggc gccaccgcgc cc #ccggccat     60 ggcttttgcc aatttccgcc gcatcctgcg cctgtctacc ttcgagaaga ga #aagtcccg    120 cgaatatgag cacgtccgcc gcgacctgga ccccaacgag gtgtgggaga tc #gtgggcga    180 gctgggcgac ggcgccttcg gcaaggttta caaggccaag aataaggaga cg #ggtgcttt    240 ggctgcggcc aaagtcattg aaaccaagag tgaggaggag ctggaggact ac #atcgtgga    300 gattgagatc ctggccacct gcgaccaccc ctacattgtg aagctcctgg ga #gcctacta    360 tcacgacggg aagctgtgga tcatgattga gttctgtcca gggggagccg tg #gacgccat    420 catgctggag ctggacagag gcctcacgga gccccagata caggtggttt gc #cgccagat    480 gctagaagcc ctcaacttcc tgcacagcaa gaggatcatc caccgagatc tg #aaagctgg    540 caacgtgctg atgaccctcg agggagacat caggctggct gactttggtg tg #tctgccaa    600 gaatctgaag actctacaga aacgagattc cttcatcggc acgccttact gg #atggcccc    660 cgaggtggtc atgtgtgaga ccatgaaaga cacgccctac gactacaaag cc #gacatctg    720 gtccctgggc atcacgctga ttgagatggc ccagatcgag ccgccacacc ac #gagctcaa    780 ccccatgcgg gtcctgctaa agatcgccaa gtcggaccct cccacgctgc tc #acgccctc    840 caagtggtct gtagagttcc gtgacttcct gaagatagcc ctggataaga ac #ccagaaac    900 ccgacccagt gccgcgcagc tgctggagca tcccttcgtc agcagcatca cc #agtaacaa    960 ggctctgcgg gagctggtgg ctgaggccaa ggccgaggtg atggaagaga tc #gaagacgg   1020 ccgggatgag ggggaagagg aggacgccgt ggatgccgcc tccaccctgg ag #aaccatac   1080 tcagaactcc tctgaggtga gtccgccaag cctcaatgct gacaagcctc tc #gaggagtc   1140 accttccacc ccgctggcac ccagccagtc tcaggacagt gtgaatgagc cc #tgcagcca   1200 gccctctggg gacagatccc tccaaaccac cagtccccca gtcgtggccc ct #ggaaatga   1260 gaacggcctg gcagtgcctg tgcccctgcg gaagtcccga cccgtgtcaa tg #gatgccag   1320 aattcaggta gcccaggaga agcaagttgc tgagcagggt ggggacctca gc #ccagcagc   1380 caacagatct caaaaggcca gccagagccg gcccaacagc agcgccctgg ag #accttggg   1440 tggggagaag ctggccaatg gcagcctgga gccacctgcc caggcagctc ca #gggccttc   1500 caagagggac tcggactgca gcagcctctg cacctctgag agcatggact at #ggtaccaa   1560 tctctccact gacctgtcgc tgaacaaaga gatgggctct ctgtccatca ag #gacccgaa   1620 actgtacaaa aaaaccctca agcggacacg caaatttgtg gtggatggtg tg #gaggtgag   1680 catcaccacc tccaagatca tcagcgaaga tgagaagaag gatgaggaga tg #agatttct   1740 caggcgccag gaactccgag agcttcggct gctccagaaa gaagagcatc gg #aaccagac   1800 ccagctgagt aacaagcatg agctgcagct ggagcaaatg cataaacgtt tt #gaacagga   1860 aatcaacgcc aagaagaagt tctttgacac ggaattagag aacctggagc gt #cagcaaaa   1920 gcagcaagtg gagaagatgg agcaagacca tgccgtgcgc cgccgggagg ag #gccaggcg   1980 gatccgcctg gagcaggatc gggactacac caggttccaa gagcagctca aa #ctgatgaa   2040 gaaagaggtg aagaacgagg tggagaagct cccccgacag cagcggaagg aa #agcatgaa   2100 gcagaagatg gaggagcaca cgcagaaaaa gcagcttctt gaccgggact tt #gtagccaa   2160 gcagaaggag gacctggagc tggccatgaa gaggctcacc accgacaaca gg #cgggagat   2220 ctgtgacaag gagcgcgagt gcctcatgaa gaagcaggag ctccttcgag ac #cgggaagc   2280 agccctgtgg gagatggaag agcaccagct gcaggagagg caccagctgg tg #aagcagca   2340 gctcaaagac cagtacttcc tccagcggca cgagctgctg cgcaagcatg ag #aaggagcg   2400 ggagcagatg cagcgctaca accagcgcat gatagagcag ctgaaggtgc gg #cagcaaca   2460 ggaaaaggcg cggctgccca agatccagag gagtgagggc aagacgcgca tg #gccatgta   2520 caagaagagc ctccacatca acggcggggg cagcgcagct gagcagcgtg ag #aagatcaa   2580 gcagttctcc cagcaggagg agaagaggca gaagtcggag cggctgcagc aa #cagcagaa   2640 acacgagaac cagatgcggg acatgctggc gcagtgcgag agcaacatga gc #gagctgca   2700 gcagctgcag aatgaaaagt gccacctcct ggtagagcac gaaacccaga aa #ctgaaggc   2760 cctggatgag agccataacc agaacctgaa ggaat        #                   #     2795 <210> SEQ ID NO 26 <211> LENGTH: 912 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 26 Met Ala Phe Ala Asn Phe Arg Arg Ile Leu Ar #g Leu Ser Thr Phe Glu   1               5  #                 10  #                 15 Lys Arg Lys Ser Arg Glu Tyr Glu His Val Ar #g Arg Asp Leu Asp Pro              20      #             25      #             30 Asn Glu Val Trp Glu Ile Val Gly Glu Leu Gl #y Asp Gly Ala Phe Gly          35          #         40          #         45 Lys Val Tyr Lys Ala Lys Asn Lys Glu Thr Gl #y Ala Leu Ala Ala Ala      50              #     55              #     60 Lys Val Ile Glu Thr Lys Ser Glu Glu Glu Le #u Glu Asp Tyr Ile Val  65                  # 70                  # 75                  # 80 Glu Ile Glu Ile Leu Ala Thr Cys Asp His Pr #o Tyr Ile Val Lys Leu                  85  #                 90  #                 95 Leu Gly Ala Tyr Tyr His Asp Gly Lys Leu Tr #p Ile Met Ile Glu Phe             100       #           105       #           110 Cys Pro Gly Gly Ala Val Asp Ala Ile Met Le #u Glu Leu Asp Arg Gly         115           #       120           #       125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Ar #g Gln Met Leu Glu Ala     130               #   135               #   140 Leu Asn Phe Leu His Ser Lys Arg Ile Ile Hi #s Arg Asp Leu Lys Ala 145                 1 #50                 1 #55                 1 #60 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Il #e Arg Leu Ala Asp Phe                 165   #               170   #               175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gl #n Lys Arg Asp Ser Phe             180       #           185       #           190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Va #l Val Met Cys Glu Thr         195           #       200           #       205 Met Lys Asp Thr Pro Tyr Asp Tyr Lys Ala As #p Ile Trp Ser Leu Gly     210               #   215               #   220 Ile Thr Leu Ile Glu Met Ala Gln Ile Glu Pr #o Pro His His Glu Leu 225                 2 #30                 2 #35                 2 #40 Asn Pro Met Arg Val Leu Leu Lys Ile Ala Ly #s Ser Asp Pro Pro Thr                 245   #               250   #               255 Leu Leu Thr Pro Ser Lys Trp Ser Val Glu Ph #e Arg Asp Phe Leu Lys             260       #           265       #           270 Ile Ala Leu Asp Lys Asn Pro Glu Thr Arg Pr #o Ser Ala Ala Gln Leu         275           #       280           #       285 Leu Glu His Pro Phe Val Ser Ser Ile Thr Se #r Asn Lys Ala Leu Arg     290               #   295               #   300 Glu Leu Val Ala Glu Ala Lys Ala Glu Val Me #t Glu Glu Ile Glu Asp 305                 3 #10                 3 #15                 3 #20 Gly Arg Asp Glu Gly Glu Glu Glu Asp Ala Va #l Asp Ala Ala Ser Thr                 325   #               330   #               335 Leu Glu Asn His Thr Gln Asn Ser Ser Glu Va #l Ser Pro Pro Ser Leu             340       #           345       #           350 Asn Ala Asp Lys Pro Leu Glu Glu Ser Pro Se #r Thr Pro Leu Ala Pro         355           #       360           #       365 Ser Gln Ser Gln Asp Ser Val Asn Glu Pro Cy #s Ser Gln Pro Ser Gly     370               #   375               #   380 Asp Arg Ser Leu Gln Thr Thr Ser Pro Pro Va #l Val Ala Pro Gly Asn 385                 3 #90                 3 #95                 4 #00 Glu Asn Gly Leu Ala Val Pro Val Pro Leu Ar #g Lys Ser Arg Pro Val                 405   #               410   #               415 Ser Met Asp Ala Arg Ile Gln Val Ala Gln Gl #u Lys Gln Val Ala Glu             420       #           425       #           430 Gln Gly Gly Asp Leu Ser Pro Ala Ala Asn Ar #g Ser Gln Lys Ala Ser         435           #       440           #       445 Gln Ser Arg Pro Asn Ser Ser Ala Leu Glu Th #r Leu Gly Gly Glu Lys     450               #   455               #   460 Leu Ala Asn Gly Ser Leu Glu Pro Pro Ala Gl #n Ala Ala Pro Gly Pro 465                 4 #70                 4 #75                 4 #80 Ser Lys Arg Asp Ser Asp Cys Ser Ser Leu Cy #s Thr Ser Glu Ser Met                 485   #               490   #               495 Asp Tyr Gly Thr Asn Leu Ser Thr Asp Leu Se #r Leu Asn Lys Glu Met             500       #           505       #           510 Gly Ser Leu Ser Ile Lys Asp Pro Lys Leu Ty #r Lys Lys Thr Leu Lys         515           #       520           #       525 Arg Thr Arg Lys Phe Val Val Asp Gly Val Gl #u Val Ser Ile Thr Thr     530               #   535               #   540 Ser Lys Ile Ile Ser Glu Asp Glu Lys Lys As #p Glu Glu Met Arg Phe 545                 5 #50                 5 #55                 5 #60 Leu Arg Arg Gln Glu Leu Arg Glu Leu Arg Le #u Leu Gln Lys Glu Glu                 565   #               570   #               575 His Arg Asn Gln Thr Gln Leu Ser Asn Lys Hi #s Glu Leu Gln Leu Glu             580       #           585       #           590 Gln Met His Lys Arg Phe Glu Gln Glu Ile As #n Ala Lys Lys Lys Phe         595           #       600           #       605 Phe Asp Thr Glu Leu Glu Asn Leu Glu Arg Gl #n Gln Lys Gln Gln Val     610               #   615               #   620 Glu Lys Met Glu Gln Asp His Ala Val Arg Ar #g Arg Glu Glu Ala Arg 625                 6 #30                 6 #35                 6 #40 Arg Ile Arg Leu Glu Gln Asp Arg Asp Tyr Th #r Arg Phe Gln Glu Gln                 645   #               650   #               655 Leu Lys Leu Met Lys Lys Glu Val Lys Asn Gl #u Val Glu Lys Leu Pro             660       #           665       #           670 Arg Gln Gln Arg Lys Glu Ser Met Lys Gln Ly #s Met Glu Glu His Thr         675           #       680           #       685 Gln Lys Lys Gln Leu Leu Asp Arg Asp Phe Va #l Ala Lys Gln Lys Glu     690               #   695               #   700 Asp Leu Glu Leu Ala Met Lys Arg Leu Thr Th #r Asp Asn Arg Arg Glu 705                 7 #10                 7 #15                 7 #20 Ile Cys Asp Lys Glu Arg Glu Cys Leu Met Ly #s Lys Gln Glu Leu Leu                 725   #               730   #               735 Arg Asp Arg Glu Ala Ala Leu Trp Glu Met Gl #u Glu His Gln Leu Gln             740       #           745       #           750 Glu Arg His Gln Leu Val Lys Gln Gln Leu Ly #s Asp Gln Tyr Phe Leu         755           #       760           #       765 Gln Arg His Glu Leu Leu Arg Lys His Glu Ly #s Glu Arg Glu Gln Met     770               #   775               #   780 Gln Arg Tyr Asn Gln Arg Met Ile Glu Gln Le #u Lys Val Arg Gln Gln 785                 7 #90                 7 #95                 8 #00 Gln Glu Lys Ala Arg Leu Pro Lys Ile Gln Ar #g Ser Glu Gly Lys Thr                 805   #               810   #               815 Arg Met Ala Met Tyr Lys Lys Ser Leu His Il #e Asn Gly Gly Gly Ser             820       #           825       #           830 Ala Ala Glu Gln Arg Glu Lys Ile Lys Gln Ph #e Ser Gln Gln Glu Glu         835           #       840           #       845 Lys Arg Gln Lys Ser Glu Arg Leu Gln Gln Gl #n Gln Lys His Glu Asn     850               #   855               #   860 Gln Met Arg Asp Met Leu Ala Gln Cys Glu Se #r Asn Met Ser Glu Leu 865                 8 #70                 8 #75                 8 #80 Gln Gln Leu Gln Asn Glu Lys Cys His Leu Le #u Val Glu His Glu Thr                 885   #               890   #               895 Gln Lys Leu Lys Ala Leu Asp Glu Ser His As #n Gln Asn Leu Lys Glu             900       #           905       #           910 <210> SEQ ID NO 27 <211> LENGTH: 3604 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 27 cgttcctggg cttcccgctc cgcaggcctg cggaggactg gcccagcaag gt #cccaggtc     60 ttccctctcc ttagcgccta agagagaggc ccagtgcggg tgaggagtcg cg #aggaagag    120 gcggaaggcg ccggaaggca ccatgttccg caagaaaaag aagaaacgcc ct #gagatctc    180 agcgccacag aacttccagc accgtgtcca cacctccttc gaccccaaag aa #ggcaagtt    240 tgtgggcctc cccccacaat ggcagaacat cctggacaca ctgcggcgcc cc #aagcccgt    300 ggtggaccct tcgcgaatca cacgggtgca gctccagccc atgaagacag tg #gtgcgggg    360 cagcgcgatg cctgtggatg gctacatctc ggggctgctc aacgacatcc ag #aagttgtc    420 agtcatcagc tccaacaccc tgcgtggccg cagccccacc agccggcggc gg #gcacagtc    480 cctggggctg ctgggggatg agcactgggc caccgaccca gacatgtacc tc #cagagccc    540 ccagtctgag cgcactgacc cccacggcct ctacctcagc tgcaacgggg gc #acaccagc    600 aggccacaag cagatgccgt ggcccgagcc acagagccca cgggtcctgc cc #aatgggct    660 ggctgcaaag gcacagtccc tgggccccgc cgagtttcag ggtgcctcgc ag #cgctgtct    720 gcagctgggt gcctgcctgc agagctcccc accaggagcc tcgcccccca cg #ggcaccaa    780 taggcatgga atgaaggctg ccaagcatgg ctctgaggag gcccggccac ag #tcctgcct    840 ggtgggctca gccacaggca ggccaggtgg ggaaggcagc cctagcccta ag #acccggga    900 gagcagcctg aagcgcaggc tattccgaag catgttcctg tccactgctg cc #acagcccc    960 tccaagcagc agcaagccag gccctccacc acagagcaag cccaactcct ct #ttccgacc   1020 gccgcagaaa gacaaccccc caagcctggt ggccaaggcc cagtccttgc cc #tcggacca   1080 gccggtgggg accttcagcc ctctgaccac ttcggatacc agcagccccc ag #aagtccct   1140 ccgcacagcc ccggccacag gccagcttcc aggccggtct tccccagcgg ga #tccccccg   1200 cacctggcac gcccagatca gcaccagcaa cctgtacctg ccccaggacc cc #acggttgc   1260 caagggtgcc ctggctggtg aggacacagg tgttgtgaca catgagcagt tc #aaggctgc   1320 gctcaggatg gtggtggacc agggtgaccc ccggctgctg ctggacagct ac #gtgaagat   1380 tggcgagggc tccaccggca tcgtctgctt ggcccgggag aagcactcgg gc #cgccaggt   1440 ggccgtcaag atgatggacc tcaggaagca gcagcgcagg gagctgctct tc #aacgaggt   1500 ggtgatcatg cgggactacc agcacttcaa cgtggtggag atgtacaaga gc #tacctggt   1560 gggcgaggag ctgtgggtgc tcatggagtt cctgcaggga ggagccctca ca #gacatcgt   1620 ctcccaagtc aggctgaatg aggagcagat tgccactgtg tgtgaggctg tg #ctgcaggc   1680 cctggcctac ctgcatgctc agggtgtcat ccaccgggac atcaagagtg ac #tccatcct   1740 gctgaccctc gatggcaggg tgaagctctc ggacttcgga ttctgtgctc ag #atcagcaa   1800 agacgtccct aagaggaagt ccctggtggg aaccccctac tggatggctc ct #gaagtgat   1860 ctccaggtct ttgtatgcca ctgaggtgga tatctggtct ctgggcatca tg #gtgattga   1920 gatggtagat ggggagccac cgtacttcag tgactcccca gtgcaagcca tg #aagaggct   1980 ccgggacagc cccccaccca agctgaaaaa ctctcacaag gtctccccag tg #ctgcgaga   2040 cttcctggag cggatgctgg tgcgggaccc ccaagagaga gccacagccc ag #gagctcct   2100 agaccacccc ttcctgctgc agacagggct acctgagtgc ctggtgcccc tg #atccagct   2160 ctaccgaaag cagacctcca cctgctgagc ccaccccaag tatgcctgcc ac #ctacgccc   2220 acaggcaggg cacactgggc agccagcctg ccggcaggac ttgcctgcct cc #tcctctca   2280 gtattctctc caaagattga aatgtgaagc cccagcccca ccctctgccc tt #cagcctac   2340 tgggccaggc cggacctgcc ccctcagtgt ctctccctcc cgagtcccca ga #tggagacc   2400 cctttctaca ggatgacccc ttgatatttg cacagggata tttctaagaa ac #gcagaggc   2460 cagcgttcct ggcctctgca gccaacacag tagaaaaggc tgctgtggtt tt #ttaaaggc   2520 agttgtccac tagtgtccta ggccactgca gagggcagac tgctggtctc ca #cagatacc   2580 tgctgttctc agctccagct tcaaacctcg agtctcgaga gggccacggg gt #ggttttta   2640 tgaccggaat cccgcttcct ccctcacgtc tgatgtcctg aaggtgcagt cc #cacctgta   2700 cagcccctcc ccgccaagaa ctgtgaatgg cctgctccag gccatggctg gg #ggcaggga   2760 gtgaggggac aatttctgag tgaaagagaa agaatggggt cggtggtgaa gg #tgctctca   2820 ctttacagaa tggagagaac atcgtgtgtg tgtgtgtgtg tgtgtgtgtg tg #tgtgtgtg   2880 tgtgtgtgtg tgtgtgtgtg tgtgtgtaag gggaggaaag ccaccttgac ag #cccaggtc   2940 cctccaggtc acccacagcc agtttcagga aggctgcccc tctctcccac ta #agttctgg   3000 cctgaaggga cctgctttct tggcctggct tccacctctc cactcctgtg tc #tacctggc   3060 cagtggagtg gtccatgcta agtctaacac tcctgggagc tcaggaggct tc #tgagcttc   3120 tcctgtactg tgcatcgtga gggccagaga caggaatgta aggattggca ac #tgtgttac   3180 ctttcaagtt tatctcaata accaggtcat cagggaccca ttgttctctt ca #gaacccta   3240 tctgggagag aaggcgaacc acctccgggt ttccatcatg tcaaggtcac ag #gcatccat   3300 gtgtgcaaac catctgcccc agctgcctcc acagactgct gtctccttgt cc #tcctcggc   3360 cctgccccac ttcagggctg ctgtgagatg gaattccagg aaagaacttc ag #gtgtctgg   3420 accctttcta tctagataat atttttagat tcttctgctc cctagtgacc ta #cctggggg   3480 caaagaaatt gcaaggactt ttttttaagg gtcagagttt tcaaaacaaa ag #catcttcc   3540 ctagaaattt ttgtgaattg tttgcacttg tgcctgtttt aaattaaatt ga #gtgttcaa   3600 agcc                  #                   #                   #           3604 <210> SEQ ID NO 28 <211> LENGTH: 2050 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 28 ggccagtggg gcgaaactgg cagctggccg gccctttaac acctacccga gg #gctgacac     60 ggaccaccca tcccggggtg cccaggggga gcctcatgac gtggccccta ac #gggccatc    120 agcggggggc ctggccatcc cccagtcctc ctcctcctcc tcccggcctc cc #acccgagc    180 ccgaggtgcc cccagccctg gagtgctggg accccacgcc tcagagcccc ag #ctggcccc    240 tccagcctgc acccccgccg cccctgctgt tcctgggccc cctggccccc gc #tcaccaca    300 gcgggagcca cagcgagtat cccatgagca gttccgggct gccctgcagc tg #gtggtgga    360 cccaggcgac ccccgctcct acctggacaa cttcatcaag attggcgagg gc #tccacggg    420 catcgtgtgc atcgccaccg tgcgcagctc gggcaagctg gtggccgtca ag #aagatgga    480 cctgcgcaag cagcagaggc gcgagctgct cttcaacgag gtggtaatca tg #agggacta    540 ccagcacgag aatgtggtgg agatgtacaa cagctacctg gtgggggacg ag #ctctgggt    600 ggtcatggag ttcctggaag gaggcgccct caccgacatc gtcacccaca cc #aggatgaa    660 cgaggagcag atcgcggccg tgtgccttgc agtgctgcag gccctgtcgg tg #ctccacgc    720 ccagggcgtc atccaccggg acatcaagag cgactcgatc ctgctgaccc at #gatggcag    780 ggtgaagctg tcagactttg ggttctgcgc ccaggtgagc aaggaagtgc cc #cgaaggaa    840 gtcgctggtc ggcacgccct actggatggc cccagagctc atctcccgcc tt #ccctacgg    900 gccagaggta gacatctggt cgctggggat aatggtgatt gagatggtgg ac #ggagagcc    960 cccctacttc aacgagccac ccctcaaagc catgaagatg attcgggaca ac #ctgccacc   1020 ccgactgaag aacctgcaca aggtgtcgcc atccctgaag ggcttcctgg ac #cgcctgct   1080 ggtgcgagac cctgcccagc gggccacggc agccgagctg ctgaagcacc ca #ttcctggc   1140 caaggcaggg ccgcctgcca gcatcgtgcc cctcatgcgc cagaaccgca cc #agatgagg   1200 cccagcgccc ttcccctcaa ccaaagagcc cccccgggtc acccccgccc ca #ctgaggcc   1260 agtagggggc caggcctccc actcctccca gcccgggaga tgctccgcgt gg #caccaccc   1320 tccttgctgg gggtagatga gaccctacta ctgaactcca gttttgatct cg #tgactttt   1380 agaaaaacac agggactcgt gggagcaagc gaggctccca ggacccccac cc #tctgggac   1440 aggccctccc ccatgttctt ctgtctccag gaagggcagc ggccctccca tc #actggaag   1500 tctgcagtgg gggtcgctgg gggtggagag aacactaaga ggtgaacatg ta #tgagtgtg   1560 tgcacgcgtg tgagtgtgca tgtgtgtgtg tgtgcaaagg tccagccacc cc #gtcctcca   1620 gcccgcaagg ggtgtctggc gccttgcctg acacccagcc ccctctcccc ct #gagccatt   1680 gtgggggtcg atcatgaatg tccgaagagt ggccttttcc cgtagccctg cg #cccccttt   1740 ctgtggctgg atggggagac aggtcagggc cccccaccct ctccagcccc tg #cagcaaat   1800 gactactgca cctggacagc ctcctctttt ctagaagtct atttatattg tc #attttata   1860 acactctagc ccctgccctt attgggggac agatggtccc tgtcctgcgg gg #tggccctg   1920 gcagaaccac tgcctgaaga accaggttcc tgcccggtca gcgcagcccc ag #cccgccca   1980 cccctgcctc gagttagttt tacaattaaa acattgtctt gttttgtgaa aa #aaaaaaaa   2040 aaaaaaaaaa                 #                   #                   #      2050 <210> SEQ ID NO 29 <211> LENGTH: 681 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 29 Met Phe Arg Lys Lys Lys Lys Lys Arg Pro Gl #u Ile Ser Ala Pro Gln   1               5  #                 10  #                 15 Asn Phe Gln His Arg Val His Thr Ser Phe As #p Pro Lys Glu Gly Lys              20      #             25      #             30 Phe Val Gly Leu Pro Pro Gln Trp Gln Asn Il #e Leu Asp Thr Leu Arg          35          #         40          #         45 Arg Pro Lys Pro Val Val Asp Pro Ser Arg Il #e Thr Arg Val Gln Leu      50              #     55              #     60 Gln Pro Met Lys Thr Val Val Arg Gly Ser Al #a Met Pro Val Asp Gly  65                  # 70                  # 75                  # 80 Tyr Ile Ser Gly Leu Leu Asn Asp Ile Gln Ly #s Leu Ser Val Ile Ser                  85  #                 90  #                 95 Ser Asn Thr Leu Arg Gly Arg Ser Pro Thr Se #r Arg Arg Arg Ala Gln             100       #           105       #           110 Ser Leu Gly Leu Leu Gly Asp Glu His Trp Al #a Thr Asp Pro Asp Met         115           #       120           #       125 Tyr Leu Gln Ser Pro Gln Ser Glu Arg Thr As #p Pro His Gly Leu Tyr     130               #   135               #   140 Leu Ser Cys Asn Gly Gly Thr Pro Ala Gly Hi #s Lys Gln Met Pro Trp 145                 1 #50                 1 #55                 1 #60 Pro Glu Pro Gln Ser Pro Arg Val Leu Pro As #n Gly Leu Ala Ala Lys                 165   #               170   #               175 Ala Gln Ser Leu Gly Pro Ala Glu Phe Gln Gl #y Ala Ser Gln Arg Cys             180       #           185       #           190 Leu Gln Leu Gly Ala Cys Leu Gln Ser Ser Pr #o Pro Gly Ala Ser Pro         195           #       200           #       205 Pro Thr Gly Thr Asn Arg His Gly Met Lys Al #a Ala Lys His Gly Ser     210               #   215               #   220 Glu Glu Ala Arg Pro Gln Ser Cys Leu Val Gl #y Ser Ala Thr Gly Arg 225                 2 #30                 2 #35                 2 #40 Pro Gly Gly Glu Gly Ser Pro Ser Pro Lys Th #r Arg Glu Ser Ser Leu                 245   #               250   #               255 Lys Arg Arg Leu Phe Arg Ser Met Phe Leu Se #r Thr Ala Ala Thr Ala             260       #           265       #           270 Pro Pro Ser Ser Ser Lys Pro Gly Pro Pro Pr #o Gln Ser Lys Pro Asn         275           #       280           #       285 Ser Ser Phe Arg Pro Pro Gln Lys Asp Asn Pr #o Pro Ser Leu Val Ala     290               #   295               #   300 Lys Ala Gln Ser Leu Pro Ser Asp Gln Pro Va #l Gly Thr Phe Ser Pro 305                 3 #10                 3 #15                 3 #20 Leu Thr Thr Ser Asp Thr Ser Ser Pro Gln Ly #s Ser Leu Arg Thr Ala                 325   #               330   #               335 Pro Ala Thr Gly Gln Leu Pro Gly Arg Ser Se #r Pro Ala Gly Ser Pro             340       #           345       #           350 Arg Thr Trp His Ala Gln Ile Ser Thr Ser As #n Leu Tyr Leu Pro Gln         355           #       360           #       365 Asp Pro Thr Val Ala Lys Gly Ala Leu Ala Gl #y Glu Asp Thr Gly Val     370               #   375               #   380 Val Thr His Glu Gln Phe Lys Ala Ala Leu Ar #g Met Val Val Asp Gln 385                 3 #90                 3 #95                 4 #00 Gly Asp Pro Arg Leu Leu Leu Asp Ser Tyr Va #l Lys Ile Gly Glu Gly                 405   #               410   #               415 Ser Thr Gly Ile Val Cys Leu Ala Arg Glu Ly #s His Ser Gly Arg Gln             420       #           425       #           430 Val Ala Val Lys Met Met Asp Leu Arg Lys Gl #n Gln Arg Arg Glu Leu         435           #       440           #       445 Leu Phe Asn Glu Val Val Ile Met Arg Asp Ty #r Gln His Phe Asn Val     450               #   455               #   460 Val Glu Met Tyr Lys Ser Tyr Leu Val Gly Gl #u Glu Leu Trp Val Leu 465                 4 #70                 4 #75                 4 #80 Met Glu Phe Leu Gln Gly Gly Ala Leu Thr As #p Ile Val Ser Gln Val                 485   #               490   #               495 Arg Leu Asn Glu Glu Gln Ile Ala Thr Val Cy #s Glu Ala Val Leu Gln             500       #           505       #           510 Ala Leu Ala Tyr Leu His Ala Gln Gly Val Il #e His Arg Asp Ile Lys         515           #       520           #       525 Ser Asp Ser Ile Leu Leu Thr Leu Asp Gly Ar #g Val Lys Leu Ser Asp     530               #   535               #   540 Phe Gly Phe Cys Ala Gln Ile Ser Lys Asp Va #l Pro Lys Arg Lys Ser 545                 5 #50                 5 #55                 5 #60 Leu Val Gly Thr Pro Tyr Trp Met Ala Pro Gl #u Val Ile Ser Arg Ser                 565   #               570   #               575 Leu Tyr Ala Thr Glu Val Asp Ile Trp Ser Le #u Gly Ile Met Val Ile             580       #           585       #           590 Glu Met Val Asp Gly Glu Pro Pro Tyr Phe Se #r Asp Ser Pro Val Gln         595           #       600           #       605 Ala Met Lys Arg Leu Arg Asp Ser Pro Pro Pr #o Lys Leu Lys Asn Ser     610               #   615               #   620 His Lys Val Ser Pro Val Leu Arg Asp Phe Le #u Glu Arg Met Leu Val 625                 6 #30                 6 #35                 6 #40 Arg Asp Pro Gln Glu Arg Ala Thr Ala Gln Gl #u Leu Leu Asp His Pro                 645   #               650   #               655 Phe Leu Leu Gln Thr Gly Leu Pro Glu Cys Le #u Val Pro Leu Ile Gln             660       #           665       #           670 Leu Tyr Arg Lys Gln Thr Ser Thr Cys         675           #       680 <210> SEQ ID NO 30 <211> LENGTH: 398 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 30 Ala Ser Gly Ala Lys Leu Ala Ala Gly Arg Pr #o Phe Asn Thr Tyr Pro   1               5  #                 10  #                 15 Arg Ala Asp Thr Asp His Pro Ser Arg Gly Al #a Gln Gly Glu Pro His              20      #             25      #             30 Asp Val Ala Pro Asn Gly Pro Ser Ala Gly Gl #y Leu Ala Ile Pro Gln          35          #         40          #         45 Ser Ser Ser Ser Ser Ser Arg Pro Pro Thr Ar #g Ala Arg Gly Ala Pro      50              #     55              #     60 Ser Pro Gly Val Leu Gly Pro His Ala Ser Gl #u Pro Gln Leu Ala Pro  65                  # 70                  # 75                  # 80 Pro Ala Cys Thr Pro Ala Ala Pro Ala Val Pr #o Gly Pro Pro Gly Pro                  85  #                 90  #                 95 Arg Ser Pro Gln Arg Glu Pro Gln Arg Val Se #r His Glu Gln Phe Arg             100       #           105       #           110 Ala Ala Leu Gln Leu Val Val Asp Pro Gly As #p Pro Arg Ser Tyr Leu         115           #       120           #       125 Asp Asn Phe Ile Lys Ile Gly Glu Gly Ser Th #r Gly Ile Val Cys Ile     130               #   135               #   140 Ala Thr Val Arg Ser Ser Gly Lys Leu Val Al #a Val Lys Lys Met Asp 145                 1 #50                 1 #55                 1 #60 Leu Arg Lys Gln Gln Arg Arg Glu Leu Leu Ph #e Asn Glu Val Val Ile                 165   #               170   #               175 Met Arg Asp Tyr Gln His Glu Asn Val Val Gl #u Met Tyr Asn Ser Tyr             180       #           185       #           190 Leu Val Gly Asp Glu Leu Trp Val Val Met Gl #u Phe Leu Glu Gly Gly         195           #       200           #       205 Ala Leu Thr Asp Ile Val Thr His Thr Arg Me #t Asn Glu Glu Gln Ile     210               #   215               #   220 Ala Ala Val Cys Leu Ala Val Leu Gln Ala Le #u Ser Val Leu His Ala 225                 2 #30                 2 #35                 2 #40 Gln Gly Val Ile His Arg Asp Ile Lys Ser As #p Ser Ile Leu Leu Thr                 245   #               250   #               255 His Asp Gly Arg Val Lys Leu Ser Asp Phe Gl #y Phe Cys Ala Gln Val             260       #           265       #           270 Ser Lys Glu Val Pro Arg Arg Lys Ser Leu Va #l Gly Thr Pro Tyr Trp         275           #       280           #       285 Met Ala Pro Glu Leu Ile Ser Arg Leu Pro Ty #r Gly Pro Glu Val Asp     290               #   295               #   300 Ile Trp Ser Leu Gly Ile Met Val Ile Glu Me #t Val Asp Gly Glu Pro 305                 3 #10                 3 #15                 3 #20 Pro Tyr Phe Asn Glu Pro Pro Leu Lys Ala Me #t Lys Met Ile Arg Asp                 325   #               330   #               335 Asn Leu Pro Pro Arg Leu Lys Asn Leu His Ly #s Val Ser Pro Ser Leu             340       #           345       #           350 Lys Gly Phe Leu Asp Arg Leu Leu Val Arg As #p Pro Ala Gln Arg Ala         355           #       360           #       365 Thr Ala Ala Glu Leu Leu Lys His Pro Phe Le #u Ala Lys Ala Gly Pro     370               #   375               #   380 Pro Ala Ser Ile Val Pro Leu Met Arg Gln As #n Arg Thr Arg 385                 3 #90                 3 #95 <210> SEQ ID NO 31 <211> LENGTH: 1001 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       murine/human SULU3 <400> SEQUENCE: 31 Met Pro Ser Thr Asn Arg Ala Gly Ser Leu Ly #s Asp Pro Glu Ile Ala   1               5  #                 10  #                 15 Glu Leu Phe Phe Lys Glu Asp Pro Glu Lys Le #u Phe Thr Asp Leu Arg              20      #             25      #             30 Glu Ile Gly His Gly Ser Phe Gly Ala Val Ty #r Phe Ala Arg Asp Val          35          #         40          #         45 Arg Thr Asn Glu Val Val Ala Ile Lys Lys Me #t Ser Tyr Ser Gly Lys      50              #     55              #     60 Gln Ser Thr Glu Lys Trp Gln Asp Ile Ile Ly #s Glu Val Lys Phe Leu  65                  # 70                  # 75                  # 80 Gln Arg Ile Lys His Pro Asn Ser Ile Glu Ty #r Lys Gly Cys Tyr Leu                  85  #                 90  #                 95 Arg Glu His Thr Ala Trp Leu Val Met Glu Ty #r Cys Leu Gly Ser Ala             100       #           105       #           110 Ser Asp Leu Leu Glu Val His Lys Lys Pro Le #u Gln Glu Val Glu Ile         115           #       120           #       125 Ala Ala Ile Thr His Gly Ala Leu Gln Gly Le #u Ala Tyr Leu His Ser     130               #   135               #   140 His Thr Met Ile His Arg Asp Ile Lys Ala Gl #y Asn Ile Leu Leu Thr 145                 1 #50                 1 #55                 1 #60 Glu Pro Gly Gln Val Lys Leu Ala Asp Phe Gl #y Ser Ala Ser Met Ala                 165   #               170   #               175 Ser Pro Ala Asn Ser Phe Val Gly Thr Pro Ty #r Trp Met Ala Pro Glu             180       #           185       #           190 Val Ile Leu Ala Met Asp Glu Gly Gln Tyr As #p Gly Lys Val Asp Val         195           #       200           #       205 Trp Ser Leu Gly Ile Thr Cys Ile Glu Leu Al #a Glu Arg Lys Pro Pro     210               #   215               #   220 Leu Phe Asn Met Asn Ala Met Ser Ala Leu Ty #r His Ile Ala Gln Asn 225                 2 #30                 2 #35                 2 #40 Glu Ser Pro Thr Leu Gln Ser Asn Glu Trp Se #r Asp Tyr Phe Arg Asn                 245   #               250   #               255 Phe Val Asp Ser Cys Leu Gln Lys Ile Pro Gl #n Asp Arg Pro Thr Ser             260       #           265       #           270 Glu Glu Leu Leu Lys His Ile Phe Val Leu Ar #g Glu Arg Pro Glu Thr         275           #       280           #       285 Val Leu Ile Asp Leu Ile Gln Arg Thr Lys As #p Ala Val Arg Glu Leu     290               #   295               #   300 Asp Asn Leu Gln Tyr Arg Lys Met Lys Lys Le #u Leu Phe Gln Glu Ala 305                 3 #10                 3 #15                 3 #20 His Asn Gly Pro Ala Val Glu Ala Gln Glu Gl #u Glu Glu Glu Gln Asp                 325   #               330   #               335 His Gly Val Gly Arg Thr Gly Thr Val Asn Se #r Val Gly Ser Asn Gln             340       #           345       #           350 Ser Ile Pro Ser Met Ser Ile Ser Ala Ser Se #r Gln Ser Ser Ser Val         355           #       360           #       365 Asn Ser Leu Pro Asp Val Ser Asp Asp Lys Se #r Glu Leu Asp Met Met     370               #   375               #   380 Glu Gly Asp His Thr Val Met Ser Asn Ser Se #r Val Ile His Leu Lys 385                 3 #90                 3 #95                 4 #00 Pro Glu Glu Glu Asn Tyr Arg Glu Glu Gly As #p Pro Arg Thr Arg Ala                 405   #               410   #               415 Ser Asp Pro Gln Ser Pro Pro Gln Val Ser Ar #g His Lys Ser His Tyr             420       #           425       #           430 Arg Asn Arg Glu His Phe Ala Thr Ile Arg Th #r Ala Ser Leu Val Thr         435           #       440           #       445 Arg Gln Met Gln Glu His Glu Gln Asp Ser Gl #u Leu Arg Glu Gln Met     450               #   455               #   460 Ser Gly Tyr Lys Arg Met Arg Arg Gln His Gl #n Lys Gln Leu Met Thr 465                 4 #70                 4 #75                 4 #80 Leu Glu Asn Lys Leu Lys Ala Glu Met Asp Gl #u His Arg Leu Arg Leu                 485   #               490   #               495 Asp Lys Asp Leu Glu Thr Gln Arg Asn Asn Ph #e Ala Ala Glu Met Glu             500       #           505       #           510 Lys Leu Ile Lys Lys His Gln Ala Ala Met Gl #u Lys Glu Ala Lys Val         515           #       520           #       525 Met Ser Asn Glu Glu Lys Lys Phe Gln Gln Hi #s Ile Gln Ala Gln Gln     530               #   535               #   540 Lys Lys Glu Leu Asn Ser Phe Leu Glu Ser Gl #n Lys Arg Glu Tyr Lys 545                 5 #50                 5 #55                 5 #60 Leu Arg Lys Glu Gln Leu Lys Glu Glu Leu As #n Glu Asn Gln Ser Thr                 565   #               570   #               575 Pro Lys Lys Glu Lys Gln Glu Trp Leu Ser Ly #s Gln Lys Glu Asn Ile             580       #           585       #           590 Gln His Phe Gln Ala Glu Glu Glu Ala Asn Le #u Leu Arg Arg Gln Arg         595           #       600           #       605 Gln Tyr Leu Glu Leu Glu Cys Arg Arg Phe Ly #s Arg Arg Met Leu Leu     610               #   615               #   620 Gly Arg His Asn Leu Glu Gln Asp Leu Val Ar #g Glu Glu Leu Asn Lys 625                 6 #30                 6 #35                 6 #40 Arg Gln Thr Gln Lys Asp Leu Glu His Ala Me #t Leu Leu Arg Gln His                 645   #               650   #               655 Glu Ser Met Gln Glu Leu Glu Phe Arg His Le #u Asn Thr Ile Gln Lys             660       #           665       #           670 Met Arg Cys Glu Leu Ile Arg Leu Gln His Gl #n Thr Glu Leu Thr Asn         675           #       680           #       685 Gln Leu Glu Tyr Asn Lys Arg Arg Glu Arg Gl #u Leu Arg Arg Lys His     690               #   695               #   700 Val Met Glu Val Arg Gln Gln Pro Lys Ser Le #u Lys Ser Lys Glu Leu 705                 7 #10                 7 #15                 7 #20 Gln Ile Lys Lys Gln Phe Gln Asp Thr Cys Ly #s Ile Gln Thr Arg Gln                 725   #               730   #               735 Tyr Lys Ala Leu Arg Asn His Leu Leu Glu Th #r Thr Pro Lys Ser Glu             740       #           745       #           750 His Lys Ala Val Leu Lys Arg Leu Lys Glu Gl #u Gln Thr Arg Lys Leu         755           #       760           #       765 Ala Ile Leu Ala Glu Gln Tyr Asp His Ser Il #e Asn Glu Met Leu Ser     770               #   775               #   780 Thr Gln Ala Leu Arg Leu Asp Glu Ala Gln Gl #u Ala Glu Cys Gln Val 785                 7 #90                 7 #95                 8 #00 Leu Lys Met Gln Leu Gln Gln Glu Leu Glu Le #u Leu Asn Ala Tyr Gln                 805   #               810   #               815 Ser Lys Ile Lys Met Gln Ala Glu Ala Gln Hi #s Asp Arg Glu Leu Arg             820       #           825       #           830 Glu Leu Glu Gln Arg Val Ser Leu Arg Arg Al #a Leu Leu Glu Gln Lys         835           #       840           #       845 Ile Glu Glu Glu Met Leu Ala Leu Gln Asn Gl #u Arg Thr Glu Arg Ile     850               #   855               #   860 Arg Ser Leu Leu Glu Arg Gln Ala Arg Glu Il #e Glu Ala Phe Asp Ser 865                 8 #70                 8 #75                 8 #80 Glu Ser Met Arg Leu Gly Phe Ser Asn Met Va #l Leu Ser Asn Leu Ser                 885   #               890   #               895 Pro Glu Ala Phe Ser His Ser Tyr Pro Gly Al #a Ser Gly Trp Ser His             900       #           905       #           910 Asn Pro Thr Gly Gly Pro Gly Pro His Trp Gl #y His Pro Met Gly Gly         915           #       920           #       925 Pro Pro Gln Ala Trp Gly His Pro Met Gln Gl #y Gly Pro Gln Pro Trp     930               #   935               #   940 Gly His Pro Ser Gly Pro Met Gln Gly Val Pr #o Arg Gly Ser Ser Met 945                 9 #50                 9 #55                 9 #60 Gly Val Arg Asn Ser Pro Gln Ala Leu Arg Ar #g Thr Ala Ser Gly Gly                 965   #               970   #               975 Arg Thr Glu Gln Gly Met Ser Arg Ser Thr Se #r Val Thr Ser Gln Ile             980       #           985       #           990 Ser Asn Gly Ser His Met Ser Tyr Thr         995           #      1000 <210> SEQ ID NO 32 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (11) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (14) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (20) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 32 ctgaattcgg ngcnttyggn aargt           #                   #               25 <210> SEQ ID NO 33 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (13) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (16) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 33 gctggatccy tcnggnggca tcca           #                   #                24 <210> SEQ ID NO 34 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (3) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (9) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (15) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 34 gcnttyggng argtntayga rgg            #                   #                23 <210> SEQ ID NO 35 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (13) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (16) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 35 gctggatccy tcnggnswca tcca           #                   #                24 <210> SEQ ID NO 36 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (9) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (15) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <220> FEATURE: <221> NAME/KEY: modified_base <222> LOCATION: (21) <223> OTHER INFORMATION: a, t, c, g, other o #r unknown <400> SEQUENCE: 36 gagttyggng argtnttyyt ngc            #                   #                23 <210> SEQ ID NO 37 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <400> SEQUENCE: 37 Gly Ala Phe Gly Lys Val   1               5 <210> SEQ ID NO 38 <211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <400> SEQUENCE: 38 Trp Met Pro Pro Glu   1               5 <210> SEQ ID NO 39 <211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <400> SEQUENCE: 39 Ala Phe Gly Glu Val Tyr Glu Gly   1               5 <210> SEQ ID NO 40 <211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <400> SEQUENCE: 40 Trp Met Ser Pro Glu   1               5 <210> SEQ ID NO 41 <211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <400> SEQUENCE: 41 Glu Phe Gly Glu Val Tyr Glu Gly   1               5 <210> SEQ ID NO 42 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 42 cacagaaacg gtcagattca c            #                   #                   #21 <210> SEQ ID NO 43 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 43 gatcagggtg acatcaaggg ac            #                   #                 22 <210> SEQ ID NO 44 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 44 ctcatctgta cacacttcat gg            #                   #                 22 <210> SEQ ID NO 45 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 45 gattcccaca ctgtagatgt c            #                   #                   #21 <210> SEQ ID NO 46 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 46 ggccctcgac tacatccacc acat           #                   #                24 <210> SEQ ID NO 47 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 47 caacgaaact aacacagcat aagg           #                   #                24 <210> SEQ ID NO 48 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 48 atggcgaacg actctcccgc gaa            #                   #                23 <210> SEQ ID NO 49 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 49 acaccaaaat caacaagttt cacctc           #                   #              26 <210> SEQ ID NO 50 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 50 agttacaagg aattccaagt tct            #                   #                23 <210> SEQ ID NO 51 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 51 atgaagagga agaaatcaaa ctg            #                   #                23 <210> SEQ ID NO 52 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 52 agatggactg tactgggagg             #                   #                   # 20 <210> SEQ ID NO 53 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 53 actttgtgca gctctgtggg             #                   #                   # 20 <210> SEQ ID NO 54 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 54 aaggttatgg atgtcacagg g            #                   #                   #21 <210> SEQ ID NO 55 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 55 ctcacaaggt tgccaacagg             #                   #                   # 20 <210> SEQ ID NO 56 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 56 agtccccacc agaaggttta c            #                   #                   #21 <210> SEQ ID NO 57 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 57 tcaggggtca gaggtcacg              #                   #                   # 19 <210> SEQ ID NO 58 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 58 cccaaaccct accacaaatt c            #                   #                   #21 <210> SEQ ID NO 59 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 59 cccccgggaa acgatgacca             #                   #                   # 20 <210> SEQ ID NO 60 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 60 agccgctgcc cctcctctac tgt            #                   #                23 <210> SEQ ID NO 61 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 61 accgcaacat cgccacctac tac            #                   #                23 <210> SEQ ID NO 62 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 62 ctcgacgtcg tggaccacc              #                   #                   # 19 <210> SEQ ID NO 63 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 63 caatgttaac ccactctatg tctc           #                   #                24 <210> SEQ ID NO 64 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 64 agtttgccga tgtttttctt ttc            #                   #                23 <210> SEQ ID NO 65 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 65 ccgccatgaa ccccggctt              #                   #                   # 19 <210> SEQ ID NO 66 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 66 cgattgccaa agaccgtgtc a            #                   #                   #21 <210> SEQ ID NO 67 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 67 agaagttgca gctgttgaga gga            #                   #                23 <210> SEQ ID NO 68 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 68 tatggcccgt gtaaggattt c            #                   #                   #21 <210> SEQ ID NO 69 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 69 gtgccagaag tgttgtgttg taa            #                   #                23 <210> SEQ ID NO 70 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 70 tattgaattg gcggaacgga ag            #                   #                 22 <210> SEQ ID NO 71 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 71 ttgttttgtg ctcattcttt ggag           #                   #                24 <210> SEQ ID NO 72 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 72 gattgctttg tgctcattct ttgg           #                   #                24 <210> SEQ ID NO 73 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 73 ttgttctaag agtgccctcc g            #                   #                   #21 <210> SEQ ID NO 74 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 74 aagaccatgc cgtgcgccg              #                   #                   # 19 <210> SEQ ID NO 75 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 75 attccttcag gttctggtta tgg            #                   #                23 <210> SEQ ID NO 76 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 76 agtccgccaa gcctcaatg              #                   #                   # 19 <210> SEQ ID NO 77 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 77 caccttcagc tgctctatca t            #                   #                   #21 <210> SEQ ID NO 78 <211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       peptide 540A <400> SEQUENCE: 78 His Gly Asp Pro Arg Pro Glu Pro Arg Pro Th #r Gln   1               5  #                 10 <210> SEQ ID NO 79 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Syntbetic       peptide 539A <400> SEQUENCE: 79 Cys Leu Asp Phe Pro Lys Glu Asp Tyr Arg   1               5  #                 10 <210> SEQ ID NO 80 <211> LENGTH: 19 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       peptide 541A <400> SEQUENCE: 80 Asp Pro Arg Thr Arg Ala Ser Asp Pro Gln Se #r Pro Pro Gln Val Ser   1               5  #                 10  #                 15 Arg His Lys <210> SEQ ID NO 81 <211> LENGTH: 14 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       peptide 542A <400> SEQUENCE: 81 Pro Ser Thr Asn Arg Ala Gly Ser Leu Lys As #p Pro Glu Cys   1               5  #                 10 <210> SEQ ID NO 82 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       peptide 554A <400> SEQUENCE: 82 Cys Leu Val Pro Leu Ile Gln Leu Tyr Arg Ly #s Gln Thr Ser Thr   1               5  #                 10  #                 15 <210> SEQ ID NO 83 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       peptide 579A <400> SEQUENCE: 83 Cys Pro Leu Met Arg Gln Asn Arg Thr Arg   1               5  #                 10 <210> SEQ ID NO 84 <211> LENGTH: 426 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 84 Met Ala His Leu Arg Gly Phe Ala Asn Gln Hi #s Ser Arg Val Asp Pro   1               5  #                 10  #                 15 Glu Glu Leu Phe Thr Lys Leu Asp Arg Ile Gl #y Lys Gly Ser Phe Gly              20      #             25      #             30 Glu Val Tyr Lys Gly Ile Asp Asn His Thr Ly #s Glu Val Val Ala Ile          35          #         40          #         45 Lys Ile Ile Asp Leu Glu Glu Ala Glu Asp Gl #u Ile Glu Asp Ile Gln      50              #     55              #     60 Gln Glu Ile Thr Val Leu Ser Gln Cys Asp Se #r Pro Tyr Ile Thr Arg  65                  # 70                  # 75                  # 80 Tyr Phe Gly Ser Tyr Leu Lys Ser Thr Lys Le #u Trp Ile Ile Met Glu                  85  #                 90  #                 95 Tyr Leu Gly Gly Gly Ser Ala Leu Asp Leu Le #u Lys Pro Gly Pro Leu             100       #           105       #           110 Glu Glu Thr Tyr Ile Ala Thr Ile Leu Arg Gl #u Ile Leu Lys Gly Leu         115           #       120           #       125 Asp Tyr Leu His Ser Glu Arg Lys Ile His Ar #g Asp Ile Lys Ala Ala     130               #   135               #   140 Asn Val Leu Leu Ser Glu Gln Gly Asp Val Ly #s Leu Ala Asp Phe Gly 145                 1 #50                 1 #55                 1 #60 Val Ala Gly Gln Leu Thr Asp Thr Gln Ile Ly #s Arg Asn Thr Phe Val                 165   #               170   #               175 Gly Thr Pro Phe Trp Met Ala Pro Glu Val Il #e Lys Gln Ser Ala Tyr             180       #           185       #           190 Asp Phe Lys Ala Asp Ile Trp Ser Leu Gly Il #e Thr Ala Ile Glu Leu         195           #       200           #       205 Ala Lys Gly Glu Pro Pro Asn Ser Asp Leu Hi #s Pro Met Arg Val Leu     210               #   215               #   220 Phe Leu Ile Pro Lys Asn Ser Pro Pro Thr Le #u Glu Gly Gln His Ser 225                 2 #30                 2 #35                 2 #40 Lys Pro Phe Lys Glu Phe Val Glu Ala Cys Le #u Asn Lys Asp Pro Arg                 245   #               250   #               255 Phe Arg Pro Thr Ala Lys Glu Leu Leu Lys Hi #s Lys Phe Ile Thr Arg             260       #           265       #           270 Tyr Thr Lys Lys Thr Ser Phe Leu Thr Glu Le #u Ile Asp Arg Tyr Lys         275           #       280           #       285 Arg Trp Lys Ser Glu Gly His Gly Glu Glu Se #r Ser Ser Glu Asp Ser     290               #   295               #   300 Asp Ile Asp Gly Glu Ala Glu Asp Gly Glu Gl #n Gly Pro Ile Trp Thr 305                 3 #10                 3 #15                 3 #20 Phe Pro Pro Thr Ile Arg Pro Ser Pro His Se #r Lys Leu His Lys Gly                 325   #               330   #               335 Thr Ala Leu His Ser Ser Gln Lys Pro Ala Gl #u Pro Val Lys Arg Gln             340       #           345       #           350 Pro Arg Ser Gln Cys Leu Ser Thr Leu Val Ar #g Pro Val Phe Gly Glu         355           #       360           #       365 Leu Lys Glu Lys His Lys Gln Ser Gly Gly Se #r Val Gly Ala Leu Glu     370               #   375               #   380 Glu Leu Glu Asn Ala Phe Ser Leu Ala Glu Gl #u Ser Cys Pro Gly Ile 385                 3 #90                 3 #95                 4 #00 Ser Asp Lys Leu Met Val His Leu Val Glu Ar #g Val Gln Arg Phe Ser                 405   #               410   #               415 His Asn Arg Asn His Leu Thr Ser Thr Arg             420       #           425 <210> SEQ ID NO 85 <211> LENGTH: 431 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 85 Met Ala His Ser Pro Val Gln Ser Gly Leu Pr #o Gly Met Gln Asn Leu   1               5  #                 10  #                 15 Lys Ala Asp Pro Glu Glu Leu Phe Thr Lys Le #u Glu Lys Ile Gly Lys              20      #             25      #             30 Gly Ser Phe Gly Glu Val Phe Lys Gly Ile As #p Asn Arg Thr Gln Lys          35          #         40          #         45 Val Val Ala Ile Lys Ile Ile Asp Leu Glu Gl #u Ala Glu Asp Glu Ile      50              #     55              #     60 Glu Asp Ile Gln Gln Glu Ile Thr Val Leu Se #r Gln Cys Asp Ser Pro  65                  # 70                  # 75                  # 80 Tyr Val Thr Lys Tyr Tyr Gly Ser Tyr Leu Ly #s Asp Thr Lys Leu Trp                  85  #                 90  #                 95 Ile Ile Met Glu Tyr Leu Gly Gly Gly Ser Al #a Leu Asp Leu Leu Glu             100       #           105       #           110 Pro Gly Pro Leu Asp Glu Thr Gln Ile Ala Th #r Ile Leu Arg Glu Ile         115           #       120           #       125 Leu Lys Gly Leu Asp Tyr Leu His Ser Glu Ly #s Lys Ile His Arg Asp     130               #   135               #   140 Ile Lys Ala Ala Asn Val Leu Leu Ser Glu Hi #s Gly Glu Val Lys Leu 145                 1 #50                 1 #55                 1 #60 Ala Asp Phe Gly Val Ala Gly Gln Leu Thr As #p Thr Gln Ile Lys Arg                 165   #               170   #               175 Asn Thr Phe Val Gly Thr Pro Phe Trp Met Al #a Pro Glu Val Ile Lys             180       #           185       #           190 Gln Ser Ala Tyr Asp Ser Lys Ala Asp Ile Tr #p Ser Leu Gly Ile Thr         195           #       200           #       205 Ala Ile Glu Leu Ala Arg Gly Glu Pro Pro Hi #s Ser Glu Leu His Pro     210               #   215               #   220 Met Lys Val Leu Phe Leu Ile Pro Lys Asn As #n Pro Pro Thr Leu Glu 225                 2 #30                 2 #35                 2 #40 Gly Asn Tyr Ser Lys Pro Leu Lys Glu Phe Va #l Glu Ala Cys Leu Asn                 245   #               250   #               255 Lys Glu Pro Ser Phe Arg Pro Thr Ala Lys Gl #u Leu Leu Lys His Lys             260       #           265       #           270 Phe Ile Leu Arg Asn Ala Lys Lys Thr Ser Ty #r Leu Thr Glu Leu Ile         275           #       280           #       285 Asp Arg Tyr Lys Arg Trp Lys Ala Glu Gln Se #r His Asp Asp Ser Ser     290               #   295               #   300 Ser Glu Asp Ser Asp Ala Glu Thr Asp Gly Gl #n Ala Ser Gly Gly Ser 305                 3 #10                 3 #15                 3 #20 Asp Ser Gly Asp Trp Ile Phe Thr Ile Arg Gl #u Lys Asp Pro Lys Asn                 325   #               330   #               335 Leu Glu Asn Gly Ala Leu Gln Pro Ser Asp Le #u Asp Arg Asn Lys Met             340       #           345       #           350 Lys Asp Ile Pro Lys Arg Pro Phe Ser Gln Cy #s Leu Ser Thr Ile Ile         355           #       360           #       365 Ser Pro Leu Phe Ala Glu Leu Lys Glu Lys Se #r Gln Ala Cys Gly Gly     370               #   375               #   380 Asn Leu Gly Ser Ile Glu Glu Leu Arg Gly Al #a Ile Tyr Leu Ala Glu 385                 3 #90                 3 #95                 4 #00 Glu Ala Cys Pro Gly Ile Ser Asp Thr Met Va #l Ala Gln Leu Val Gln                 405   #               410   #               415 Arg Leu Gln Arg Tyr Ser Leu Ser Gly Gly Gl #y Thr Ser Ser His             420       #           425       #           430 <210> SEQ ID NO 86 <211> LENGTH: 443 <212> TYPE: PRT <213> ORGANISM: Caenorhabditis elegans <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (389)..(390) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 86 Met Thr Thr Thr Ser Ser Asp Glu Leu Pro Ar #g Gln Ala Asp Asp Asp   1               5  #                 10  #                 15 Ser Met Lys Trp Asp Arg Ile Tyr Ile Gln Ly #s Leu Asp Pro Glu Val              20      #             25      #             30 Ile Phe Thr Lys Gln Glu Arg Ile Gly Arg Gl #y Ser Phe Gly Glu Val          35          #         40          #         45 Tyr Lys Gly Ile Asp Asn Arg Thr Gly Arg Va #l Val Ala Ile Lys Ile      50              #     55              #     60 Ile Asp Leu Glu Gln Ala Glu Asp Glu Ile Gl #u Asp Ile Gln Gln Glu  65                  # 70                  # 75                  # 80 Ile Gln Val Leu Ser Gln Cys Asp Ser Gln Ty #r Val Thr Lys Tyr Phe                  85  #                 90  #                 95 Gly Ser Phe Leu Lys Gly Ser Lys Leu Trp Il #e Ile Met Glu Tyr Leu             100       #           105       #           110 Gly Gly Gly Ser Ala Leu Asp Leu Thr Lys Se #r Gly Lys Leu Asp Glu         115           #       120           #       125 Ser His Ile Ala Val Ile Leu Arg Glu Ile Le #u Lys Gly Leu Glu Tyr     130               #   135               #   140 Leu His Ser Glu Arg Lys Ile His Arg Asp Il #e Lys Ala Ala Asn Val 145                 1 #50                 1 #55                 1 #60 Leu Val Ser Glu His Gly Asp Val Lys Val Al #a Asp Phe Gly Val Ala                 165   #               170   #               175 Gly Gln Leu Thr Glu Thr Val Lys Lys Arg Il #e Thr Phe Val Gly Ser             180       #           185       #           190 Pro Phe Trp Met Ala Pro Glu Leu Ile Lys Gl #n Ser Ser Tyr Asp Tyr         195           #       200           #       205 Lys Ala Asp Ile Trp Ser Leu Gly Ile Thr Al #a Ile Glu Leu Ala Asn     210               #   215               #   220 Gly Glu Pro Pro His Ser Asp Leu His Pro Me #t Arg Val Leu Phe Leu 225                 2 #30                 2 #35                 2 #40 Ile Pro Lys Asn Pro Pro Pro Val Leu Gln Gl #y Ser Gln Trp Ser Lys                 245   #               250   #               255 Pro Phe Lys Glu Phe Val Glu Met Cys Leu As #n Lys Asp Pro Glu Asn             260       #           265       #           270 Arg Pro Ser Ala Ser Thr Leu Leu Lys His Gl #n Phe Ile Lys Arg Ala         275           #       280           #       285 Lys Lys Asn Ser Ile Leu Val Asp Leu Ile Gl #u Arg Ala Ala Glu Tyr     290               #   295               #   300 Arg Leu Arg Thr Gly Val Ser Ser Asp Ser As #p Leu Asp Glu Asp Ser 305                 3 #10                 3 #15                 3 #20 Asp Gly Gly Gly Gly Thr Ser Lys Trp Asp Ty #r Pro Thr Val Arg Gly                 325   #               330   #               335 Pro Arg Val Ser Ala Asp Asp Asp Gly Thr Va #l Arg Gln Arg Thr Asp             340       #           345       #           350 Arg Pro Arg Ala Gln Val Asp Arg Arg Ser Pr #o Ser Gly Ser Pro Gly         355           #       360           #       365 Gly Thr Ile Val Arg Gly Ser Pro Gln Val Al #a Ala Val Ala Glu Gln     370               #   375               #   380 Leu Arg Asn Ser Xaa Xaa Ala Leu Asp Gln Le #u Arg His Val Phe Arg 385                 3 #90                 3 #95                 4 #00 Asp Val Glu Asp Ser Cys Pro Gly Ile Cys As #n Glu Leu Ile Glu Glu                 405   #               410   #               415 Leu Met Gln Arg Ile Ala Val Pro Gln Val Se #r Gln Ser Asp Leu Asp             420       #           425       #           430 Ala Ala Ile Arg Arg Leu Thr Thr Pro Pro Se #r         435           #       440 <210> SEQ ID NO 87 <211> LENGTH: 275 <212> TYPE: PRT <213> ORGANISM: Saccharomyces pombe <400> SEQUENCE: 87 Leu Leu Tyr Arg Asn Phe Val Lys Ile Gly Gl #n Gly Ala Ser Gly Asp   1               5  #                 10  #                 15 Val Tyr Ser Ala Arg Gln Val Gly Thr Asn Le #u Ser Val Ala Ile Lys              20      #             25      #             30 Lys Met Asn Ile Asn Gln Gln Pro Lys Lys Gl #u Phe Ile Val Asn Glu          35          #         40          #         45 Ile Leu Val Met Lys Ser His His His Lys As #n Ile Val Asn Phe Ile      50              #     55              #     60 Asp Thr Phe Phe Tyr Lys Ser Glu Leu Trp Me #t Val Met Glu Tyr Met  65                  # 70                  # 75                  # 80 Arg Gly Gly Ser Leu Thr Glu Val Val Thr As #n Asn Thr Leu Ser Glu                  85  #                 90  #                 95 Gly Gln Ile Ala Ala Ile Cys Lys Glu Thr Le #u Glu Gly Leu Gln His             100       #           105       #           110 Leu His Glu Asn Gly Ile Val His Arg Asp Il #e Lys Ser Asp Asn Ile         115           #       120           #       125 Leu Leu Ser Leu Gln Gly Asp Ile Lys Leu Th #r Asp Phe Gly Phe Cys     130               #   135               #   140 Ala Gln Ile Asp Ser Asn Met Thr Lys Arg Th #r Thr Met Val Gly Thr 145                 1 #50                 1 #55                 1 #60 Pro Tyr Trp Met Ala Pro Glu Val Val Thr Ar #g Lys Glu Tyr Gly Phe                 165   #               170   #               175 Lys Val Asp Val Trp Ser Leu Gly Ile Met Al #a Ile Glu Met Val Glu             180       #           185       #           190 Gly Glu Pro Pro Tyr Leu Asn Glu Asn Pro Le #u Arg Ala Leu Tyr Leu         195           #       200           #       205 Ile Ala Thr Ile Gly Thr Pro Lys Ile Ser Ar #g Pro Glu Leu Leu Ser     210               #   215               #   220 Ser Val Phe His Asp Phe Leu Ser Lys Ser Le #u Thr Val Asn Pro Lys 225                 2 #30                 2 #35                 2 #40 Gln Arg Pro Ser Ser Gly Glu Leu Leu Arg Hi #s Pro Phe Leu Lys Gln                 245   #               250   #               255 Ala Val Pro Val Ser Ser Leu Ile Pro Leu Il #e Lys Ser Ile His His             260       #           265       #           270 Ser Gly Lys         275 <210> SEQ ID NO 88 <211> LENGTH: 1109 <212> TYPE: PRT <213> ORGANISM: Caenorhabditis elegans <400> SEQUENCE: 88 Met Ser Ser Ser Gly Leu Asp Glu Ile Asp Le #u Asn Ser Leu Arg Asp   1               5  #                 10  #                 15 Pro Ala Gly Ile Phe Glu Leu Ile Glu Val Va #l Gly Asn Gly Thr Tyr              20      #             25      #             30 Gly Gln Val Tyr Lys Gly Arg His Val Lys Th #r Ala Gln Leu Ala Ala          35          #         40          #         45 Ile Lys Ile Met Asn Ile Asn Glu Asp Glu Gl #u Asp Glu Ile Lys Leu      50              #     55              #     60 Glu Ile Asn Met Leu Lys Lys His Ser His Hi #s Arg Asn Val Ala Thr  65                  # 70                  # 75                  # 80 Tyr Tyr Gly Ala Phe Ile Lys Lys Leu Pro Se #r Ser Thr Gly Lys His                  85  #                 90  #                 95 Asp Gln Leu Trp Leu Val Met Glu Phe Cys Gl #y Ser Gly Ser Ile Thr             100       #           105       #           110 Asp Leu Val Lys Asn Thr Lys Gly Gly Ser Le #u Lys Glu Glu Trp Ile         115           #       120           #       125 Ala Tyr Ile Cys Arg Glu Ile Leu Arg Gly Le #u Tyr His Leu His Gln     130               #   135               #   140 Ser Lys Val Ile His Arg Asp Ile Lys Gly Gl #n Asn Val Leu Leu Thr 145                 1 #50                 1 #55                 1 #60 Asp Ser Ala Glu Val Lys Leu Val Asp Phe Gl #y Val Ser Ala Gln Leu                 165   #               170   #               175 Asp Lys Thr Val Gly Arg Arg Asn Thr Phe Il #e Gly Thr Pro Tyr Trp             180       #           185       #           190 Met Ala Pro Glu Val Ile Ala Cys Asp Glu Se #r Pro Glu Ala Thr Tyr         195           #       200           #       205 Asp Ser Arg Ser Asp Leu Trp Ser Leu Gly Il #e Thr Ala Leu Glu Met     210               #   215               #   220 Ala Glu Gly His Pro Pro Leu Cys Asp Met Hi #s Pro Met Arg Ala Leu 225                 2 #30                 2 #35                 2 #40 Phe Leu Ile Pro Arg Asn Pro Pro Pro Lys Le #u Lys Arg Asn Lys Lys                 245   #               250   #               255 Trp Thr Lys Lys Phe Glu Thr Phe Ile Glu Th #r Val Leu Val Lys Asp             260       #           265       #           270 Tyr His Gln Arg Pro Tyr Thr Gly Ala Leu Le #u Arg His Pro Phe Ile         275           #       280           #       285 Lys Glu Gln Pro His Glu Gln Thr Ile Arg Hi #s Ser Ile Lys Glu His     290               #   295               #   300 Ile Asp Arg Asn Arg Arg Val Lys Lys Asp As #p Ala Asp Tyr Glu Tyr 305                 3 #10                 3 #15                 3 #20 Ser Gly Ser Glu Asp Asp Glu Pro Ser Pro As #n Asn Arg Asp Asp Ser                 325   #               330   #               335 Glu Ser Ser Ser Met Ile Pro Met Asp Asn Th #r Leu Arg Lys Gly Phe             340       #           345       #           350 Gln Lys Leu Gln Glu Ser Ser Arg Gly Phe Al #a Glu Pro Gly Ala Gln         355           #       360           #       365 Gln Leu Arg Arg Leu Pro Gln Gln Pro Ala Pr #o Ala Pro Phe Gln Tyr     370               #   375               #   380 Gln Gln Ser Arg Tyr Val Glu Pro Arg Arg Gl #u Ser Ser Glu Val Lys 385                 3 #90                 3 #95                 4 #00 Leu Arg Ala Val Ser Ser Arg Gly Ala Ala As #p Gly Pro Arg His Ser                 405   #               410   #               415 Pro Ala Ser Arg Pro Arg Pro Arg Ser Pro Gl #n Gln Ser His Pro Ala             420       #           425       #           430 Ala Pro His Leu Ala Asp Leu Ala Asn Tyr Gl #u Lys Arg Arg Arg Ser         435           #       440           #       445 Glu Arg Glu Glu Arg Arg Glu Arg Glu Arg Gl #n Ala His His Ala Met     450               #   455               #   460 Pro Ile Ala Arg Val Ser Ala Ser Val Pro Al #a Pro Gln Gln Ser Arg 465                 4 #70                 4 #75                 4 #80 Lys Met Ser Glu Pro Leu Leu Ile Thr His Va #l Lys Pro Glu Asp Leu                 485   #               490   #               495 Asp Val Leu Ala Ser Glu Leu Ser Lys Met Gl #y Gly His His Asn Gly             500       #           505       #           510 Arg Ser Arg Glu Glu Ser Met Ser Pro Pro Pr #o Pro Ala Pro Pro Pro         515           #       520           #       525 Arg Glu Ala Ser Ile Ser Ser Ile Thr Asp Th #r Ile Asp Val Gly Glu     530               #   535               #   540 Leu Asp Asn Gly Ala Asp Ala Glu Trp Asp As #p Leu Lys Asp Ile Met 545                 5 #50                 5 #55                 5 #60 Met Asn Gly Glu Gly Thr Leu Arg Gly Pro As #n Lys Pro Leu Pro Pro                 565   #               570   #               575 Thr Pro Thr Asp Gly Glu Asn Thr Leu Val Se #r Asp Val Arg Arg Asn             580       #           585       #           590 Gly Asn Gly Asn Ser Gly His Gly Ala Tyr Ly #s Gly Lys Lys Ile Pro         595           #       600           #       605 Glu Ile Arg Pro Gly Ile Ile Ser Leu Asp As #p Asp Asp Ser Asp Ser     610               #   615               #   620 Asp Asn Glu Glu Gly Asn Glu Pro Leu Met Ph #e Lys Pro Ile Val Arg 625                 6 #30                 6 #35                 6 #40 Cys Pro Phe Ser Ile Phe Phe Trp Phe Leu Se #r Ala Asn Val Ile His                 645   #               650   #               655 Ser Val Asp Gly Ser Ile Pro Leu Val Lys Hi #s Leu Ile Trp Phe Gln             660       #           665       #           670 Asn Ala Ser Ser Ser Arg Gly Ala Leu Pro As #p Leu Leu Pro Lys Ser         675           #       680           #       685 Pro Asp Leu Arg Arg Gln Ile Asn Asp Gln Th #r Arg Gln Met Ser Asp     690               #   695               #   700 Asp Arg Ala Asp Glu Gln Pro Asn Gly Phe Gl #n Asn Ser Asp Ser Arg 705                 7 #10                 7 #15                 7 #20 Ser Ser Ile Gln His Ser Phe Ser Asn Arg As #p Arg Glu Lys Ser Phe                 725   #               730   #               735 Val Gly Tyr Phe Gly Gly Gly Ala Gly Ala Gl #y Gly Gly Thr Val Asn             740       #           745       #           750 Arg Pro Gly Arg Pro Gln Asp Ile Asn Gln Va #l Gln Val Asn Val Thr         755           #       760           #       765 Pro Asn Ser Asn Gly Thr Pro Ala Glu Asn As #p Ala Pro Glu Ile Arg     770               #   775               #   780 Lys Tyr Lys Lys Lys Phe Ser Gly Glu Ile Le #u Cys Ala Ala Leu Trp 785                 7 #90                 7 #95                 8 #00 Gly Val Asn Leu Leu Ile Gly Thr Asp Ser Gl #y Leu Met Leu Leu Asp                 805   #               810   #               815 Arg Ser Gly Gln Gly Lys Val Tyr Pro Leu Il #e Ser Arg Arg Arg Phe             820       #           825       #           830 Asp Gln Met Thr Val Leu Glu Gly Gln Asn Il #e Leu Ala Thr Ile Ser         835           #       840           #       845 Gly Arg Lys Arg Arg Ile Arg Val Tyr Tyr Le #u Ser Trp Leu Arg Gln     850               #   855               #   860 Lys Ile Leu Arg Thr Glu Gly Ala Gly Ser Al #a Asn Thr Thr Glu Lys 865                 8 #70                 8 #75                 8 #80 Arg Asn Gly Trp Val Asn Val Gly Asp Leu Gl #n Gly Ala Ile His Phe                 885   #               890   #               895 Lys Ile Val Arg Tyr Glu Arg Ile Lys Phe Le #u Val Val Gly Leu Glu             900       #           905       #           910 Ser Ser Ile Glu Ile Tyr Ala Trp Ala Pro Ly #s Pro Tyr His Lys Phe         915           #       920           #       925 Met Ser Phe Lys Ser Phe Gly Ser Leu Ser Hi #s Val Pro Leu Ile Val     930               #   935               #   940 Asp Leu Thr Val Glu Asp Asn Ala Arg Leu Ly #s Val Leu Tyr Gly Ser 945                 9 #50                 9 #55                 9 #60 Thr Gly Gly Phe His Ala Ile Asp Leu Asp Se #r Ala Ala Val Tyr Asp                 965   #               970   #               975 Ile Tyr Thr Pro Ala Gln Ser Gly Gln Thr Th #r Thr Pro His Cys Ile             980       #           985       #           990 Val Val Leu Pro Asn Ser Asn Gly Met Gln Le #u Leu Leu Cys Tyr Asp         995           #      1000            #     1005 Asn Glu Gly Val Tyr Val Asn Thr Tyr Gly Ar #g Met Thr Lys Asn Val    1010               #  1015                # 1020 Val Leu Gln Trp Gly Glu Met Pro Ser Ser Va #l Ala Tyr Ile Ser Thr 1025               1030  #               1035   #              1040 Gly Gln Ile Met Gly Trp Gly Asn Lys Ala Il #e Glu Ile Arg Ser Val                1045   #              1050    #             1055 Asp Thr Gly His Leu Asp Gly Val Phe Met Hi #s Lys Lys Ala Gln Lys            1060       #          1065        #         1070 Leu Lys Phe Leu Cys Glu Arg Asn Asp Lys Va #l Phe Phe Ser Ser Ala        1075           #      1080            #     1085 Lys Gly Gly Gly Ser Cys Gln Ile Tyr Phe Me #t Thr Leu Asn Lys Pro    1090               #  1095                # 1100 Gly Leu Thr Asn Trp 1105 <210> SEQ ID NO 89 <211> LENGTH: 1233 <212> TYPE: PRT <213> ORGANISM: Murine sp. <400> SEQUENCE: 89 Met Ala Asn Asp Ser Pro Ala Lys Ser Leu Va #l Asp Ile Asp Leu Ser   1               5  #                 10  #                 15 Ser Leu Arg Asp Pro Ala Gly Ile Phe Glu Le #u Val Glu Val Val Gly              20      #             25      #             30 Asn Gly Thr Tyr Gly Gln Val Tyr Lys Gly Ar #g His Val Lys Thr Val          35          #         40          #         45 Thr Ala Ala Ile Lys Val Met Asp Val Thr Gl #u Asp Glu Glu Glu Glu      50              #     55              #     60 Ile Thr Leu Glu Ile Asn Met Leu Lys Lys Ty #r Ser His His Arg Asn  65                  # 70                  # 75                  # 80 Ile Ala Thr Tyr Tyr Gly Ala Phe Ile Lys Ly #s Ser Pro Pro Gly His                  85  #                 90  #                 95 Asp Asp Gln Leu Trp Leu Val Met Glu Phe Cy #s Gly Ala Gly Ser Ile             100       #           105       #           110 Thr Asp Leu Val Lys Asn Thr Lys Gly Asn Th #r Leu Lys Glu Asp Trp         115           #       120           #       125 Ile Ala Tyr Ile Ser Arg Glu Ile Leu Arg Gl #y Leu Ala His Leu His     130               #   135               #   140 Ile His His Val Ile His Arg Asp Ile Lys Gl #y Gln Asn Val Leu Leu 145                 1 #50                 1 #55                 1 #60 Thr Glu Asn Ala Glu Val Lys Leu Val Asp Ph #e Gly Val Ser Ala Gln                 165   #               170   #               175 Leu Asp Arg Thr Val Gly Arg Arg Asn Thr Ph #e Ile Gly Thr Pro Tyr             180       #           185       #           190 Trp Met Ala Pro Glu Val Ile Ala Cys Asp Gl #u Asn Pro Asp Ala Thr         195           #       200           #       205 Tyr Asp Tyr Arg Ser Asp Leu Trp Ser Cys Gl #y Ile Thr Ala Ile Glu     210               #   215               #   220 Met Ala Glu Gly Gly Pro Pro Leu Cys Asp Me #t His Pro Met Arg Ala 225                 2 #30                 2 #35                 2 #40 Leu Phe Leu Ile Pro Arg Asn Pro Pro Pro Ar #g Leu Lys Ser Lys Lys                 245   #               250   #               255 Trp Ser Lys Lys Phe Phe Ser Phe Ile Glu Gl #y Cys Leu Val Lys Asn             260       #           265       #           270 Tyr Met Gln Arg Pro Ser Thr Glu Gln Leu Le #u Lys His Pro Phe Ile         275           #       280           #       285 Arg Asp Gln Pro Asn Glu Arg Gln Val Arg Il #e Gln Leu Lys Asp His     290               #   295               #   300 Ile Asp Arg Thr Arg Lys Lys Arg Gly Glu Ly #s Asp Glu Thr Glu Tyr 305                 3 #10                 3 #15                 3 #20 Glu Tyr Ser Gly Ser Glu Glu Glu Glu Glu Gl #u Val Pro Glu Gln Glu                 325   #               330   #               335 Gly Glu Pro Ser Ser Ile Val Asn Val Pro Gl #y Glu Ser Thr Leu Arg             340       #           345       #           350 Arg Asp Phe Leu Arg Leu Gln Gln Glu Asn Ly #s Glu Arg Ser Glu Ala         355           #       360           #       365 Leu Arg Arg Gln Gln Leu Leu Gln Glu Gln Gl #n Leu Arg Glu Gln Glu     370               #   375               #   380 Glu Tyr Lys Arg Gln Leu Leu Ala Glu Arg Gl #n Lys Arg Ile Glu Gln 385                 3 #90                 3 #95                 4 #00 Gln Lys Glu Gln Arg Arg Arg Leu Glu Glu Gl #n Gln Arg Arg Glu Arg                 405   #               410   #               415 Glu Ala Arg Arg Gln Gln Glu Arg Glu Gln Ar #g Arg Arg Glu Gln Glu             420       #           425       #           430 Glu Lys Arg Arg Leu Glu Glu Leu Glu Arg Ar #g Arg Lys Glu Glu Glu         435           #       440           #       445 Glu Arg Arg Arg Ala Glu Glu Glu Lys Arg Ar #g Val Glu Arg Glu Gln     450               #   455               #   460 Glu Tyr Ile Arg Arg Gln Leu Glu Glu Glu Gl #n Arg His Leu Glu Ile 465                 4 #70                 4 #75                 4 #80 Leu Gln Gln Gln Leu Leu Gln Glu Gln Ala Me #t Leu Leu His Asp His                 485   #               490   #               495 Arg Arg Pro His Ala Gln Gln Gln Pro Pro Pr #o Pro Gln Gln Gln Asp             500       #           505       #           510 Arg Ser Lys Pro Ser Phe His Ala Pro Glu Pr #o Lys Pro His Tyr Asp         515           #       520           #       525 Pro Ala Asp Arg Ala Arg Glu Val Gln Trp Se #r His Leu Ala Ser Leu     530               #   535               #   540 Lys Asn Asn Val Ser Pro Val Ser Arg Ser Hi #s Ser Phe Ser Asp Pro 545                 5 #50                 5 #55                 5 #60 Ser Pro Lys Phe Ala His His His Leu Arg Se #r Gln Asp Pro Cys Pro                 565   #               570   #               575 Pro Ser Arg Ser Glu Gly Leu Ser Gln Ser Se #r Asp Ser Lys Ser Glu             580       #           585       #           590 Val Pro Glu Pro Thr Gln Lys Ala Trp Ser Ar #g Ser Asp Ser Asp Glu         595           #       600           #       605 Val Pro Pro Arg Val Pro Val Arg Thr Thr Se #r Arg Ser Pro Val Leu     610               #   615               #   620 Ser Arg Arg Asp Ser Pro Leu Gln Gly Gly Gl #y Gln Gln Asn Ser Gln 625                 6 #30                 6 #35                 6 #40 Ala Gly Gln Arg Asn Ser Thr Ser Ser Ile Gl #u Pro Arg Leu Leu Trp                 645   #               650   #               655 Glu Arg Val Glu Lys Leu Val Pro Arg Pro Gl #y Ser Gly Ser Ser Ser             660       #           665       #           670 Gly Ser Ser Asn Ser Gly Ser Gln Pro Gly Se #r His Pro Gly Ser Gln         675           #       680           #       685 Ser Gly Ser Gly Glu Arg Phe Arg Val Arg Se #r Ser Ser Lys Ser Glu     690               #   695               #   700 Gly Ser Pro Ser Pro Arg Gln Glu Ser Ala Al #a Lys Lys Pro Asp Asp 705                 7 #10                 7 #15                 7 #20 Lys Lys Glu Val Phe Arg Ser Leu Lys Pro Al #a Gly Glu Val Asp Leu                 725   #               730   #               735 Thr Ala Leu Ala Lys Glu Leu Arg Ala Val Gl #u Asp Val Arg Pro Pro             740       #           745       #           750 His Lys Val Thr Asp Tyr Ser Ser Ser Ser Gl #u Glu Ser Gly Thr Thr         755           #       760           #       765 Asp Glu Glu Glu Glu Asp Val Glu Gln Glu Gl #y Ala Asp Asp Ser Thr     770               #   775               #   780 Ser Gly Pro Glu Asp Thr Arg Ala Ala Ser Se #r Pro Asn Leu Ser Asn 785                 7 #90                 7 #95                 8 #00 Gly Glu Thr Glu Ser Val Lys Thr Met Ile Va #l His Asp Asp Val Glu                 805   #               810   #               815 Ser Glu Pro Ala Met Thr Pro Ser Lys Glu Gl #y Thr Leu Ile Val Arg             820       #           825       #           830 Gln Thr Gln Ser Ala Ser Ser Thr Leu Gln Ly #s His Lys Ser Ser Ser         835           #       840           #       845 Ser Phe Thr Pro Phe Ile Asp Pro Arg Leu Le #u Gln Ile Ser Pro Ser     850               #   855               #   860 Ser Gly Thr Thr Val Thr Ser Val Val Gly Ph #e Ser Cys Asp Gly Leu 865                 8 #70                 8 #75                 8 #80 Arg Pro Glu Ala Ile Arg Gln Asp Pro Thr Ar #g Lys Gly Ser Val Val                 885   #               890   #               895 Asn Val Asn Pro Thr Asn Thr Arg Pro Gln Se #r Asp Thr Pro Glu Ile             900       #           905       #           910 Arg Lys Tyr Lys Lys Arg Phe Asn Ser Glu Il #e Leu Cys Ala Ala Leu         915           #       920           #       925 Trp Gly Val Asn Leu Leu Val Gly Thr Glu Se #r Gly Leu Met Leu Leu     930               #   935               #   940 Asp Arg Ser Gly Gln Gly Lys Val Tyr Pro Le #u Ile Ser Arg Arg Arg 945                 9 #50                 9 #55                 9 #60 Phe Gln Gln Met Asp Val Leu Glu Gly Leu As #n Val Leu Val Thr Ile                 965   #               970   #               975 Ser Gly Lys Lys Asp Lys Leu Arg Val Tyr Ty #r Leu Ser Trp Leu Arg             980       #           985       #           990 Asn Lys Ile Leu His Asn Asp Pro Glu Val Gl #u Lys Lys Gln Gly Trp         995           #      1000            #     1005 Thr Thr Val Gly Asp Leu Glu Gly Cys Val Hi #s Tyr Lys Val Val Lys    1010               #  1015                # 1020 Tyr Glu Arg Ile Lys Phe Leu Val Ile Ala Le #u Lys Ser Ser Val Glu 1025               1030  #               1035   #              1040 Val Tyr Ala Trp Ala Pro Lys Pro Tyr His Ly #s Phe Met Ala Phe Lys                1045   #              1050    #             1055 Ser Phe Gly Glu Leu Leu His Lys Pro Leu Le #u Val Asp Leu Thr Val            1060       #          1065        #         1070 Glu Glu Gly Gln Arg Leu Lys Val Ile Tyr Gl #y Ser Cys Ala Gly Phe        1075           #      1080            #     1085 His Ala Val Asp Val Asp Ser Gly Ser Val Ty #r Asp Ile Tyr Leu Pro    1090               #  1095                # 1100 Thr His Ile Gln Cys Ser Ile Lys Pro His Al #a Ile Ile Ile Leu Pro 1105               1110  #               1115   #              1120 Asn Thr Asp Gly Met Glu Leu Leu Val Cys Ty #r Glu Asp Glu Gly Val                1125   #              1130    #             1135 Tyr Val Asn Thr Tyr Gly Arg Ile Thr Lys As #p Val Val Leu Gln Trp            1140       #          1145        #         1150 Gly Glu Met Pro Thr Ser Val Ala Tyr Ile Ar #g Ser Asn Gln Thr Met        1155           #      1160            #     1165 Gly Trp Gly Glu Lys Ala Ile Glu Ile Arg Se #r Val Glu Thr Gly His    1170               #  1175                # 1180 Leu Asp Gly Val Phe Met His Lys Arg Ala Gl #n Arg Leu Lys Phe Leu 1185               1190  #               1195   #              1200 Cys Gly Arg Asn Asp Lys Val Phe Phe Ser Se #r Val Arg Ser Gly Gly                1205   #              1210    #             1215 Ser Ser Gln Val Tyr Phe Met Thr Leu Gly Ar #g Thr Ser Leu Leu Ser            1220       #          1225        #         1230 Trp <210> SEQ ID NO 90 <211> LENGTH: 982 <212> TYPE: PRT <213> ORGANISM: Caenorhabditis elegans <400> SEQUENCE: 90 Met Ala Pro Ala Val Leu Asp Lys Pro Gly Va #l Ile Lys Asp Pro Ser   1               5  #                 10  #                 15 Ile Ala Ala Leu Phe Ser Asn Lys Asp Pro Gl #u Gln Arg Tyr Gln Asp              20      #             25      #             30 Leu Arg Glu Ile Gly His Gly Ser Phe Gly Al #a Val Tyr Phe Ala Tyr          35          #         40          #         45 Asp Lys Lys Asn Glu Gln Thr Val Ala Ile Ly #s Lys Met Asn Phe Ser      50              #     55              #     60 Gly Lys Gln Ala Val Glu Lys Trp Asn Asp Il #e Leu Lys Glu Val Ser  65                  # 70                  # 75                  # 80 Phe Leu Asn Thr Val Val His Pro His Ile Va #l Asp Tyr Lys Ala Cys                  85  #                 90  #                 95 Phe Leu Lys Asp Thr Thr Cys Trp Leu Val Me #t Glu Tyr Cys Ile Gly             100       #           105       #           110 Ser Ala Ala Asp Ile Val Asp Val Leu Arg Ly #s Gly Met Arg Glu Val         115           #       120           #       125 Glu Ile Ala Ala Ile Cys Ser Gln Thr Leu As #p Ala Leu Arg Tyr Leu     130               #   135               #   140 His Ser Leu Lys Arg Ile His Arg Asp Ile Ly #s Ala Gly Asn Ile Leu 145                 1 #50                 1 #55                 1 #60 Leu Ser Asp His Ala Ile Val Lys Leu Ala As #p Phe Gly Ser Ala Ser                 165   #               170   #               175 Leu Val Asp Pro Ala Gln Thr Phe Ile Gly Th #r Pro Phe Phe Met Ala             180       #           185       #           190 Pro Glu Val Ile Leu Ala Met Asp Glu Gly Hi #s Tyr Thr Asp Arg Ala         195           #       200           #       205 Asp Ile Trp Ser Leu Gly Ile Thr Cys Ile Gl #u Leu Ala Glu Arg Arg     210               #   215               #   220 Pro Pro Leu Phe Ser Met Asn Ala Met Ser Al #a Leu Tyr His Ile Ala 225                 2 #30                 2 #35                 2 #40 Gln Asn Asp Pro Pro Thr Leu Ser Pro Ile As #p Thr Ser Glu Gln Pro                 245   #               250   #               255 Glu Trp Ser Leu Glu Phe Val Gln Phe Ile As #p Lys Cys Leu Arg Lys             260       #           265       #           270 Pro Ala Glu Glu Arg Met Ser Ala Glu Glu Cy #s Phe Arg His Pro Phe         275           #       280           #       285 Ile Gln Arg Ser Arg Pro Ser Asp Thr Ile Gl #n Glu Leu Ile Gln Arg     290               #   295               #   300 Thr Lys Asn Met Val Leu Glu Leu Asp Asn Ph #e Gln Tyr Lys Lys Met 305                 3 #10                 3 #15                 3 #20 Arg Lys Leu Met Tyr Leu Asp Glu Thr Glu Gl #y Lys Glu Gly Ser Glu                 325   #               330   #               335 Gly Asn Gly Ala Ser Asp Asp Leu Asp Phe Hi #s Gly Asn Glu Ala Asn             340       #           345       #           350 Ser Ile Gly Arg Ala Gly Asp Ser Ala Ser Se #r Arg Ser Ala Ser Leu         355           #       360           #       365 Thr Ser Phe Arg Ser Met Gln Ser Ser Gly Gl #y Ala Gly Leu Leu Val     370               #   375               #   380 Ser Thr Asn Thr Thr Gly Ala Met Asp Asn Va #l His Gly Ser Ser Gly 385                 3 #90                 3 #95                 4 #00 Tyr Gly Asn Gly Ser Ser Ser Thr Thr Ser Se #r Ala Arg Arg Arg Pro                 405   #               410   #               415 Pro Ile Pro Ser Gln Met Leu Ser Ser Thr Se #r Thr Ser Gly Val Gly             420       #           425       #           430 Thr Met Pro Ser His Gly Ser Val Gly Ala Se #r Ile Thr Ala Ile Ala         435           #       440           #       445 Val Asn Pro Thr Pro Ser Pro Ser Glu Pro Il #e Pro Thr Ser Gln Pro     450               #   455               #   460 Thr Ser Lys Ser Glu Ser Ser Ser Ile Leu Gl #u Thr Ala His Asp Asp 465                 4 #70                 4 #75                 4 #80 Pro Leu Asp Thr Ser Ile Arg Ala Pro Val Ly #s Asp Leu His Met Pro                 485   #               490   #               495 His Arg Ala Val Lys Glu Arg Ile Ala Thr Le #u Gln Asn His Lys Phe             500       #           505       #           510 Ala Thr Leu Arg Ser Gln Arg Ile Ile Asn Gl #n Glu Gln Glu Glu Tyr         515           #       520           #       525 Thr Lys Glu Asn Asn Met Tyr Glu Gln Met Se #r Lys Tyr Lys His Leu     530               #   535               #   540 Arg Gln Ala His His Lys Glu Leu Gln Gln Ph #e Glu Glu Arg Cys Ala 545                 5 #50                 5 #55                 5 #60 Leu Asp Arg Glu Gln Leu Arg Val Lys Met As #p Arg Glu Leu Glu Gln                 565   #               570   #               575 Leu Thr Thr Thr Tyr Ser Lys Glu Lys Met Ar #g Val Arg Cys Ser Gln             580       #           585       #           590 Asn Asn Glu Leu Asp Lys Arg Lys Lys Asp Il #e Glu Asp Gly Glu Lys         595           #       600           #       605 Lys Met Lys Lys Thr Lys Asn Ser Gln Asn Gl #n Gln Gln Met Lys Leu     610               #   615               #   620 Tyr Ser Ala Gln Gln Leu Lys Glu Tyr Lys Ty #r Asn Lys Glu Ala Gln 625                 6 #30                 6 #35                 6 #40 Lys Thr Arg Leu Arg Ser Leu Asn Met Pro Ar #g Ser Thr Tyr Glu Asn                 645   #               650   #               655 Ala Met Lys Glu Val Lys Ala Asp Leu Asn Ar #g Val Lys Asp Ala Arg             660       #           665       #           670 Glu Asn Asp Phe Asp Glu Lys Leu Arg Ala Gl #u Leu Glu Asp Glu Ile         675           #       680           #       685 Val Arg Tyr Arg Arg Gln Gln Leu Ser Asn Le #u His Gln Leu Glu Glu     690               #   695               #   700 Gln Leu Asp Asp Glu Asp Val Asn Val Gln Gl #u Arg Gln Met Asp Thr 705                 7 #10                 7 #15                 7 #20 Arg His Gly Leu Leu Ser Lys Gln His Glu Me #t Thr Arg Asp Leu Glu                 725   #               730   #               735 Ile Gln His Leu Asn Glu Leu His Ala Met Ly #s Lys Arg His Leu Glu             740       #           745       #           750 Thr Gln His Glu Ala Glu Ser Ala Ser Gln As #n Glu Tyr Thr Gln Arg         755           #       760           #       765 Gln Gln Asp Glu Leu Arg Lys Lys His Ala Me #t Gln Ser Arg Gln Gln     770               #   775               #   780 Pro Arg Asp Leu Lys Ile Gln Glu Ala Gln Il #e Arg Lys Gln Tyr Arg 785                 7 #90                 7 #95                 8 #00 Gln Val Val Lys Thr Gln Thr Arg Gln Phe Ly #s Leu Tyr Leu Thr Gln                 805   #               810   #               815 Met Val Gln Val Val Pro Lys Asp Glu Gln Ly #s Glu Leu Thr Ser Arg             820       #           825       #           830 Leu Lys Gln Asp Gln Met Gln Lys Val Ala Le #u Leu Ala Ser Gln Tyr         835           #       840           #       845 Glu Ser Gln Ile Lys Lys Met Val Gln Asp Ly #s Thr Val Lys Leu Glu     850               #   855               #   860 Ser Trp Gln Glu Asp Glu Gln Arg Val Leu Se #r Glu Lys Leu Glu Lys 865                 8 #70                 8 #75                 8 #80 Glu Leu Glu Glu Leu Ile Ala Tyr Gln Lys Ly #s Thr Arg Ala Thr Leu                 885   #               890   #               895 Glu Glu Gln Ile Lys Lys Glu Arg Thr Ala Le #u Glu Glu Arg Ile Gly             900       #           905       #           910 Thr Arg Arg Ala Met Leu Glu Gln Lys Ile Il #e Glu Glu Arg Glu Gln         915           #       920           #       925 Met Gly Glu Met Arg Arg Leu Lys Lys Glu Gl #n Ile Arg Asp Arg His     930               #   935               #   940 Ser Gln Glu Arg His Arg Leu Glu Asn His Ph #e Val Arg Thr Gly Ser 945                 9 #50                 9 #55                 9 #60 Thr Ser Arg Ser Ser Gly Gly Ile Ala Pro Gl #y Val Gly Asn Ser Ser                 965   #               970   #               975 Ser Ile Gln Met Ala Met             980 <210> SEQ ID NO 91 <211> LENGTH: 842 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 91 Leu Arg Pro Ala Ala Asp Ile Leu Arg Arg As #n Pro Gln Gln Asp Tyr   1               5  #                 10  #                 15 Glu Leu Val Gln Arg Val Gly Ser Gly Thr Ty #r Gly Asp Val Tyr Lys              20      #             25      #             30 Ala Arg Asn Val His Thr Gly Glu Leu Ala Al #a Val Lys Ile Ile Lys          35          #         40          #         45 Leu Glu Pro Gly Asp Asp Phe Ser Leu Ile Gl #n Gln Glu Ile Phe Met      50              #     55              #     60 Val Lys Glu Cys Lys His Cys Asn Ile Val Al #a Tyr Phe Gly Ser Tyr  65                  # 70                  # 75                  # 80 Leu Ser Arg Glu Lys Leu Trp Ile Cys Met Gl #u Tyr Cys Gly Gly Gly                  85  #                 90  #                 95 Ser Leu Gln Asp Ile Tyr His Val Thr Gly Pr #o Leu Ser Glu Leu Gln             100       #           105       #           110 Ile Ala Tyr Val Cys Arg Glu Thr Leu Gln Gl #y Leu Ala Tyr Leu His         115           #       120           #       125 Thr Lys Gly Lys Met His Arg Asp Ile Lys Gl #y Ala Asn Ile Leu Leu     130               #   135               #   140 Thr Asp His Gly Asp Val Lys Leu Ala Asp Ph #e Gly Val Ala Ala Lys 145                 1 #50                 1 #55                 1 #60 Ile Thr Ala Thr Ile Ala Lys Arg Lys Ser Ph #e Ile Gly Thr Pro Tyr                 165   #               170   #               175 Trp Met Ala Pro Glu Val Ala Ala Val Glu Ly #s Asn Gly Gly Tyr Asn             180       #           185       #           190 Gln Leu Cys Asp Ile Trp Ala Val Gly Ile Th #r Ala Ile Glu Leu Gly         195           #       200           #       205 Glu Leu Gln Pro Pro Met Phe Asp Leu His Pr #o Met Arg Ala Leu Phe     210               #   215               #   220 Leu Met Ser Lys Ser Asn Phe Gln Pro Pro Ly #s Leu Lys Asp Lys Thr 225                 2 #30                 2 #35                 2 #40 Lys Trp Ser Ser Thr Phe His Asn Phe Val Ly #s Ile Ala Leu Thr Lys                 245   #               250   #               255 Asn Pro Lys Lys Arg Pro Thr Ala Glu Arg Le #u Leu Thr His Thr Phe             260       #           265       #           270 Val Ala Gln Pro Gly Leu Ser Arg Ala Leu Al #a Val Glu Leu Leu Asp         275           #       280           #       285 Lys Val Asn Asn Pro Asp Asn His Ala His Ty #r Thr Glu Ala Asp Asp     290               #   295               #   300 Asp Asp Phe Glu Pro His Ala Ile Ile Arg Hi #s Thr Ile Arg Ser Thr 305                 3 #10                 3 #15                 3 #20 Asn Arg Asn Ala Arg Ala Glu Arg Thr Ala Se #r Glu Ile Asn Phe Asp                 325   #               330   #               335 Lys Leu Gln Phe Glu Pro Pro Leu Arg Lys Gl #u Thr Glu Ala Arg Asp             340       #           345       #           350 Glu Met Gly Leu Ser Ser Asp Pro Asn Phe Me #t Leu Gln Trp Asn Pro         355           #       360           #       365 Phe Val Asp Gly Ala Asn Thr Gly Lys Ser Th #r Ser Lys Arg Ala Ile     370               #   375               #   380 Pro Pro Pro Leu Pro Pro Lys Pro Arg Ile Se #r Ser Tyr Pro Glu Asp 385                 3 #90                 3 #95                 4 #00 Asn Phe Pro Asp Glu Glu Lys Ala Ser Thr Il #e Lys His Cys Pro Asp                 405   #               410   #               415 Ser Glu Ser Arg Ala Pro Gln Ile Leu Arg Ar #g Gln Ser Ser Pro Ser             420       #           425       #           430 Cys Gly Pro Val Ala Glu Thr Ser Ser Ile Gl #y Asn Gly Asp Gly Ile         435           #       440           #       445 Ser Lys Leu Met Ser Glu Asn Thr Glu Gly Se #r Ala Gln Ala Pro Gln     450               #   455               #   460 Leu Pro Arg Lys Asn Asp Lys Arg Asp Phe Pr #o Lys Pro Ala Ile Asn 465                 4 #70                 4 #75                 4 #80 Gly Leu Pro Pro Thr Pro Lys Val Leu Met Gl #y Ala Cys Phe Ser Lys                 485   #               490   #               495 Val Phe Asp Gly Cys Pro Leu Lys Ile Asn Cy #s Ala Thr Ser Trp Ile             500       #           505       #           510 His Pro Asp Thr Lys Asp Gln Tyr Ile Ile Ph #e Gly Thr Glu Asp Gly         515           #       520           #       525 Ile Tyr Thr Leu Asn Leu Asn Glu Leu His Gl #u Ala Thr Met Glu Gln     530               #   535               #   540 Leu Phe Pro Arg Lys Cys Thr Trp Leu Tyr Va #l Ile Asn Asn Thr Leu 545                 5 #50                 5 #55                 5 #60 Met Ser Leu Ser Glu Gly Lys Thr Phe Gln Le #u Tyr Ser His Asn Leu                 565   #               570   #               575 Ile Ala Leu Phe Glu His Ala Lys Lys Pro Gl #y Leu Ala Ala His Ile             580       #           585       #           590 Gln Thr His Arg Phe Pro Asp Arg Ile Leu Pr #o Arg Lys Phe Ala Leu         595           #       600           #       605 Thr Thr Lys Ile Pro Asp Thr Lys Gly Cys Hi #s Lys Cys Cys Ile Val     610               #   615               #   620 Arg Asn Pro Tyr Thr Gly His Lys Tyr Leu Cy #s Gly Ala Leu Gln Ser 625                 6 #30                 6 #35                 6 #40 Gly Ile Val Leu Leu Gln Trp Tyr Glu Pro Me #t Gln Lys Phe Met Leu                 645   #               650   #               655 Ile Lys His Phe Asp Phe Pro Leu Pro Ser Pr #o Leu Asn Val Phe Glu             660       #           665       #           670 Met Leu Val Ile Pro Glu Gln Glu Tyr Pro Me #t Val Cys Val Ala Ile         675           #       680           #       685 Ser Lys Gly Thr Glu Ser Asn Gln Val Val Gl #n Phe Glu Thr Ile Asn     690               #   695               #   700 Leu Asn Ser Ala Ser Ser Trp Phe Thr Glu Il #e Gly Ala Gly Ser Gln 705                 7 #10                 7 #15                 7 #20 Gln Leu Asp Ser Ile His Val Thr Gln Leu Gl #u Arg Asp Thr Val Leu                 725   #               730   #               735 Val Cys Leu Asp Lys Phe Val Lys Ile Val As #n Leu Gln Gly Lys Leu             740       #           745       #           750 Lys Ser Ser Lys Lys Leu Ala Ser Glu Leu Se #r Phe Asp Phe Arg Ile         755           #       760           #       765 Glu Ser Val Val Cys Leu Gln Asp Ser Val Le #u Ala Phe Trp Lys His     770               #   775               #   780 Gly Met Gln Gly Lys Ser Phe Lys Ser Asp Gl #u Val Thr Gln Glu Ile 785                 7 #90                 7 #95                 8 #00 Ser Asp Glu Thr Arg Val Phe Arg Leu Leu Gl #y Ser Asp Arg Val Val                 805   #               810   #               815 Val Leu Glu Ser Arg Pro Thr Glu Asn Pro Th #r Ala His Ser Asn Leu             820       #           825       #           830 Tyr Ile Leu Ala Gly His Glu Asn Ser Tyr         835           #       840 <210> SEQ ID NO 92 <211> LENGTH: 911 <212> TYPE: PRT <213> ORGANISM: Murine sp. <400> SEQUENCE: 92 Met Ala Phe Ala Asn Phe Arg Arg Ile Leu Ar #g Leu Ser Thr Phe Glu   1               5  #                 10  #                 15 Lys Arg Lys Ser Arg Glu Tyr Glu His Val Ar #g Arg Asp Leu Asp Pro              20      #             25      #             30 Asn Asp Val Trp Glu Ile Val Gly Glu Leu Gl #y Asp Gly Ala Phe Gly          35          #         40          #         45 Lys Val Tyr Lys Ala Lys Asn Lys Glu Thr Gl #y Ala Leu Ala Ala Ala      50              #     55              #     60 Lys Val Ile Glu Thr Lys Ser Glu Glu Glu Le #u Glu Asp Tyr Ile Val  65                  # 70                  # 75                  # 80 Glu Ile Glu Ile Leu Ala Thr Cys Asp His Pr #o Tyr Ile Val Lys Leu                  85  #                 90  #                 95 Leu Gly Ala Tyr Tyr Tyr Asp Gly Lys Leu Tr #p Ile Met Ile Glu Phe             100       #           105       #           110 Cys Pro Gly Gly Ala Val Asp Ala Ile Met Le #u Glu Leu Asp Arg Gly         115           #       120           #       125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Ar #g Gln Met Leu Glu Ala     130               #   135               #   140 Leu Asn Phe Leu His Gly Lys Arg Ile Ile Hi #s Arg Asp Leu Lys Ala 145                 1 #50                 1 #55                 1 #60 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Il #e Arg Leu Ala Asp Phe                 165   #               170   #               175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gl #n Lys Arg Asp Ser Phe             180       #           185       #           190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Va #l Val Leu Cys Glu Thr         195           #       200           #       205 Met Lys Asp Ala Pro Tyr Asp Tyr Lys Ala As #p Ile Trp Ser Leu Gly     210               #   215               #   220 Ile Thr Leu Ile Glu Met Ala Gln Ile Glu Pr #o Pro His His Glu Leu 225                 2 #30                 2 #35                 2 #40 Asn Pro Met Arg Val Leu Leu Lys Ile Ala Ly #s Ser Asp Pro Pro Thr                 245   #               250   #               255 Leu Leu Thr Pro Ser Lys Trp Ser Val Glu Ph #e Arg Asp Phe Leu Lys             260       #           265       #           270 Ile Ala Leu Asp Lys Asn Pro Glu Thr Arg Pr #o Ser Ala Ala Gln Leu         275           #       280           #       285 Leu Gln His Pro Phe Val Ser Arg Val Thr Se #r Asn Lys Ala Leu Arg     290               #   295               #   300 Glu Leu Val Ala Glu Ala Lys Ala Glu Val Me #t Glu Glu Ile Glu Asp 305                 3 #10                 3 #15                 3 #20 Gly Arg Glu Asp Gly Glu Glu Glu Asp Ala Va #l Asp Ala Val Pro Pro                 325   #               330   #               335 Leu Val Asn His Thr Gln Asp Ser Ala Asn Va #l Thr Gln Pro Ser Leu             340       #           345       #           350 Asp Ser Asn Lys Leu Leu Gln Asp Ser Ser Th #r Pro Leu Pro Pro Ser         355           #       360           #       365 Gln Pro Gln Glu Pro Val Asn Gly Pro Cys Se #r Gln Pro Ser Gly Asp     370               #   375               #   380 Gly Pro Leu Gln Thr Thr Ser Pro Ala Asp Gl #y Leu Ser Lys Asn Asp 385                 3 #90                 3 #95                 4 #00 Asn Asp Leu Lys Val Pro Val Pro Leu Arg Ly #s Ser Arg Pro Leu Ser                 405   #               410   #               415 Met Asp Ala Arg Ile Gln Met Asp Glu Glu Ly #s Gln Ile Pro Asp Gln             420       #           425       #           430 Asp Glu Asn Pro Ser Pro Ala Ala Ser Lys Se #r Gln Lys Ala Asn Gln         435           #       440           #       445 Ser Arg Pro Asn Ser Ser Ala Leu Glu Thr Le #u Gly Gly Glu Ala Leu     450               #   455               #   460 Thr Asn Gly Gly Leu Glu Leu Pro Ser Ser Va #l Thr Pro Ser His Ser 465                 4 #70                 4 #75                 4 #80 Lys Arg Ala Ser Asp Cys Ser Asn Leu Ser Th #r Ser Glu Ser Met Asp                 485   #               490   #               495 Tyr Gly Thr Ser Leu Ser Ala Asp Leu Ser Le #u Asn Lys Glu Thr Gly             500       #           505       #           510 Ser Leu Ser Leu Lys Gly Ser Lys Leu His As #n Lys Thr Leu Lys Arg         515           #       520           #       525 Thr Arg Arg Phe Val Val Asp Gly Val Glu Va #l Ser Ile Thr Thr Ser     530               #   535               #   540 Lys Ile Ile Ser Glu Asp Glu Lys Lys Asp Gl #u Glu Met Arg Phe Leu 545                 5 #50                 5 #55                 5 #60 Arg Arg Gln Glu Leu Arg Glu Leu Arg Leu Le #u Gln Lys Glu Glu His                 565   #               570   #               575 Arg Asn Gln Thr Gln Leu Ser Ser Lys His Gl #u Leu Gln Leu Glu Gln             580       #           585       #           590 Met His Lys Arg Phe Glu Gln Glu Ile Asn Al #a Lys Lys Lys Phe Tyr         595           #       600           #       605 Asp Val Glu Leu Glu Asn Leu Glu Arg Gln Gl #n Lys Gln Gln Val Glu     610               #   615               #   620 Lys Met Glu Gln Asp His Ser Val Arg Arg Ly #s Glu Glu Ala Lys Arg 625                 6 #30                 6 #35                 6 #40 Ile Arg Leu Glu Gln Asp Arg Asp Tyr Ala Ly #s Phe Gln Glu Gln Leu                 645   #               650   #               655 Lys Gln Met Lys Lys Glu Val Lys Ser Glu Va #l Glu Lys Leu Pro Arg             660       #           665       #           670 Gln Gln Arg Lys Glu Ser Met Lys Gln Lys Me #t Glu Glu His Ser Gln         675           #       680           #       685 Lys Lys Gln Arg Leu Asp Arg Asp Phe Val Al #a Lys Gln Lys Glu Asp     690               #   695               #   700 Leu Glu Leu Ala Met Arg Lys Leu Thr Thr Gl #u Asn Arg Arg Glu Ile 705                 7 #10                 7 #15                 7 #20 Cys Asp Lys Glu Arg Asp Cys Leu Ser Lys Ly #s Gln Glu Leu Leu Arg                 725   #               730   #               735 Asp Arg Glu Ala Ala Leu Trp Glu Met Glu Gl #u His Gln Leu Gln Glu             740       #           745       #           750 Arg His Gln Leu Val Lys Gln Gln Leu Lys As #p Gln Tyr Phe Leu Gln         755           #       760           #       765 Arg His Asp Leu Leu Arg Lys His Glu Lys Gl #u Arg Glu Gln Met Gln     770               #   775               #   780 Arg Tyr Asn Gln Arg Met Met Glu Gln Leu Ly #s Val Arg Gln Gln Gln 785                 7 #90                 7 #95                 8 #00 Glu Lys Ala Arg Leu Pro Lys Ile Gln Arg Se #r Asp Gly Glu Thr Arg                 805   #               810   #               815 Met Ala Met Tyr Lys Lys Ser Leu His Ile As #n Gly Ala Gly Ser Ala             820       #           825       #           830 Ser Glu Gln Arg Glu Lys Ile Lys Gln Phe Se #r Gln Gln Glu Glu Lys         835           #       840           #       845 Arg Gln Lys Ala Glu Arg Leu Gln Gln Gln Gl #n Lys His Glu His Gln     850               #   855               #   860 Met Arg Asp Met Val Ala Gln Cys Glu Ser As #n Met Ser Glu Leu Gln 865                 8 #70                 8 #75                 8 #80 Gln Leu Gln Asn Glu Lys Cys Tyr Leu Leu Va #l Glu His Glu Thr Gln                 885   #               890   #               895 Lys Leu Lys Ala Leu Asp Glu Ser His Asn Gl #n Ser Leu Lys Glu             900       #           905       #           910 <210> SEQ ID NO 93 <211> LENGTH: 545 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 93 Met Ser Asn Asn Gly Leu Asp Ile Gln Asp Ly #s Pro Pro Ala Pro Pro   1               5  #                 10  #                 15 Met Arg Asn Thr Ser Thr Met Ile Gly Ala Gl #y Ser Lys Asp Ala Gly              20      #             25      #             30 Thr Leu Asn His Gly Ser Lys Pro Leu Pro Pr #o Asn Pro Glu Glu Lys          35          #         40          #         45 Lys Lys Lys Asp Arg Phe Tyr Arg Ser Ile Le #u Pro Gly Asp Lys Thr      50              #     55              #     60 Asn Lys Lys Lys Glu Lys Glu Arg Pro Glu Il #e Ser Leu Pro Ser Asp  65                  # 70                  # 75                  # 80 Phe Glu His Thr Ile His Val Gly Phe Asp Al #a Val Thr Gly Glu Phe                  85  #                 90  #                 95 Thr Gly Met Pro Glu Gln Trp Ala Arg Leu Le #u Gln Thr Ser Asn Ile             100       #           105       #           110 Thr Lys Ser Glu Gln Lys Lys Asn Pro Gln Al #a Val Leu Asp Val Leu         115           #       120           #       125 Glu Phe Tyr Asn Ser Lys Lys Thr Ser Asn Se #r Gln Lys Tyr Met Ser     130               #   135               #   140 Phe Thr Asp Lys Ser Ala Glu Asp Tyr Asn Se #r Ser Asn Ala Leu Asn 145                 1 #50                 1 #55                 1 #60 Val Lys Ala Val Ser Glu Thr Pro Ala Val Pr #o Pro Val Ser Glu Asp                 165   #               170   #               175 Glu Asp Asp Asp Asp Asp Asp Ala Thr Pro Pr #o Pro Val Ile Ala Pro             180       #           185       #           190 Arg Pro Glu His Thr Lys Ser Val Tyr Thr Ar #g Ser Val Ile Glu Pro         195           #       200           #       205 Leu Pro Val Thr Pro Thr Arg Asp Val Ala Th #r Ser Pro Ile Ser Pro     210               #   215               #   220 Thr Glu Asn Asn Thr Thr Pro Pro Asp Ala Le #u Thr Leu Asn Thr Glu 225                 2 #30                 2 #35                 2 #40 Lys Gln Lys Lys Lys Pro Lys Met Ser Asp Gl #u Glu Ile Leu Glu Lys                 245   #               250   #               255 Leu Arg Ser Ile Val Ser Val Gly Asp Pro Ly #s Lys Lys Tyr Thr Arg             260       #           265       #           270 Phe Glu Lys Ile Gly Gln Gly Ala Ser Gly Th #r Val Tyr Thr Ala Met         275           #       280           #       285 Asp Val Ala Thr Gly Gln Glu Val Ala Ile Ly #s Gln Met Asn Leu Gln     290               #   295               #   300 Gln Gln Pro Lys Lys Glu Leu Ile Ile Asn Gl #u Ile Leu Val Met Arg 305                 3 #10                 3 #15                 3 #20 Glu Asn Lys Asn Pro Asn Ile Val Asn Tyr Le #u Asp Ser Tyr Leu Val                 325   #               330   #               335 Gly Asp Glu Leu Trp Val Val Met Glu Tyr Le #u Ala Gly Gly Ser Leu             340       #           345       #           350 Thr Asp Val Val Thr Glu Thr Cys Met Asp Gl #u Gly Gln Ile Ala Ala         355           #       360           #       365 Val Cys Arg Glu Cys Leu Gln Ala Leu Glu Se #r Leu His Ser Asn Gln     370               #   375               #   380 Val Ile His Arg Asp Ile Lys Ser Asp Asn Il #e Leu Leu Gly Met Asp 385                 3 #90                 3 #95                 4 #00 Gly Ser Val Lys Leu Thr Asp Phe Gly Phe Cy #s Ala Gln Ile Thr Pro                 405   #               410   #               415 Glu Gln Ser Lys Arg Ser Thr Met Val Gly Th #r Pro Tyr Trp Met Ala             420       #           425       #           430 Pro Glu Val Val Thr Arg Lys Ala Tyr Gly Pr #o Lys Val Asp Ile Trp         435           #       440           #       445 Ser Leu Gly Ile Met Ala Ile Glu Met Ile Gl #u Gly Glu Pro Pro Tyr     450               #   455               #   460 Leu Asn Glu Asn Pro Leu Arg Ala Leu Tyr Le #u Ile Ala Thr Asn Gly 465                 4 #70                 4 #75                 4 #80 Thr Pro Glu Leu Gln Asn Pro Glu Lys Leu Se #r Ala Ile Phe Arg Asp                 485   #               490   #               495 Phe Leu Asn Arg Cys Leu Glu Met Asp Val Gl #u Lys Arg Gly Ser Ala             500       #           505       #           510 Lys Glu Leu Leu Gln His Gln Phe Leu Lys Il #e Ala Lys Pro Leu Ser         515           #       520           #       525 Ser Leu Thr Pro Leu Ile Ala Ala Ala Lys Gl #u Ala Thr Lys Asn Asn     530               #   535               #   540 His 545 <210> SEQ ID NO 94 <211> LENGTH: 506 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 94 Met Glu Glu Thr Gln Gln Lys Ser Asn Leu Gl #u Leu Leu Ser Ala Asn   1               5  #                 10  #                 15 His Ser Leu Lys Pro Leu Pro Ser Val Pro Gl #u Glu Lys Lys Pro Arg              20      #             25      #             30 His Lys Ile Ile Ser Ile Phe Ser Gly Thr Gl #u Lys Gly Ser Lys Lys          35          #         40          #         45 Lys Glu Lys Glu Arg Pro Glu Ile Ser Pro Pr #o Ser Asp Phe Glu His      50              #     55              #     60 Thr Ile His Val Gly Phe Asp Thr Val Thr Gl #y Glu Phe Thr Gly Met  65                  # 70                  # 75                  # 80 Pro Glu Gln Trp Ala Arg Leu Leu Gln Thr Se #r Asn Ile Thr Lys Leu                  85  #                 90  #                 95 Glu Gln Lys Lys Asn Pro Gln Ala Val Leu As #p Val Leu Lys Phe Tyr             100       #           105       #           110 Asp Ser Asn Thr Val Lys Gln Lys Tyr Leu Se #r Phe Thr Pro Pro Glu         115           #       120           #       125 Lys Asp Gly Phe Pro Ser Gly Thr Pro Ala Le #u Asn Ala Lys Gly Thr     130               #   135               #   140 Glu Ala Pro Ala Val Val Thr Glu Glu Glu As #p Asp Asp Glu Glu Thr 145                 1 #50                 1 #55                 1 #60 Ala Pro Pro Val Ile Ala Pro Arg Pro Asp Hi #s Thr Lys Ser Ile Tyr                 165   #               170   #               175 Thr Arg Ser Val Ile Asp Pro Val Pro Ala Pr #o Val Gly Asp Ser His             180       #           185       #           190 Val Asp Gly Ala Ala Lys Ser Leu Asp Lys Gl #n Lys Lys Lys Thr Lys         195           #       200           #       205 Met Thr Asp Glu Glu Ile Met Glu Lys Leu Ar #g Thr Ile Val Ser Ile     210               #   215               #   220 Gly Asp Pro Lys Lys Lys Tyr Thr Arg Tyr Gl #u Lys Ile Gly Gln Gly 225                 2 #30                 2 #35                 2 #40 Ala Ser Gly Thr Val Phe Thr Ala Thr Asp Va #l Ala Leu Gly Gln Glu                 245   #               250   #               255 Val Ala Ile Lys Gln Ile Asn Leu Gln Lys Gl #n Pro Lys Lys Glu Leu             260       #           265       #           270 Ile Ile Asn Glu Ile Leu Val Met Lys Glu Le #u Lys Asn Pro Asn Ile         275           #       280           #       285 Val Asn Phe Leu Asp Ser Tyr Leu Val Gly As #p Glu Leu Phe Val Val     290               #   295               #   300 Met Glu Tyr Leu Ala Gly Arg Ser Leu Thr As #p Val Val Thr Glu Thr 305                 3 #10                 3 #15                 3 #20 Cys Met Asp Glu Ala Gln Ile Ala Ala Val Cy #s Arg Glu Cys Leu Gln                 325   #               330   #               335 Ala Leu Glu Phe Leu His Ala Asn Gln Val Il #e His Arg Asp Ile Lys             340       #           345       #           350 Ser Asp Asn Val Leu Leu Gly Met Glu Gly Se #r Val Lys Leu Thr Asp         355           #       360           #       365 Phe Gly Phe Cys Ala Gln Ile Thr Pro Glu Gl #n Ser Lys Arg Ser Thr     370               #   375               #   380 Met Val Gly Thr Pro Tyr Trp Met Ala Pro Gl #u Val Val Thr Arg Lys 385                 3 #90                 3 #95                 4 #00 Ala Tyr Gly Pro Lys Val Asp Ile Trp Ser Le #u Gly Ile Met Ala Ile                 405   #               410   #               415 Glu Met Val Glu Gly Glu Pro Pro Tyr Leu As #n Glu Asn Pro Leu Arg             420       #           425       #           430 Ala Leu Tyr Leu Ile Ala Thr Asn Gly Thr Pr #o Glu Leu Gln Asn Pro         435           #       440           #       445 Glu Lys Leu Ser Pro Ile Phe Arg Asp Phe Le #u Asn Arg Cys Leu Glu     450               #   455               #   460 Met Asp Val Glu Lys Arg Gly Ser Ala Lys Gl #u Leu Leu Gln His Pro 465                 4 #70                 4 #75                 4 #80 Phe Leu Lys Leu Ala Lys Pro Leu Ser Ser Le #u Thr Pro Leu Ile Met                 485   #               490   #               495 Ala Ala Lys Glu Ala Met Lys Ser Asn Arg             500       #           505 <210> SEQ ID NO 95 <211> LENGTH: 544 <212> TYPE: PRT <213> ORGANISM: Murine sp. <400> SEQUENCE: 95 Met Ser Asp Ser Leu Asp Asn Glu Glu Lys Pr #o Pro Ala Pro Pro Leu   1               5  #                 10  #                 15 Arg Met Asn Ser Asn Asn Arg Asp Ser Ser Al #a Leu Asn His Ser Ser              20      #             25      #             30 Lys Pro Leu Pro Met Ala Pro Glu Glu Lys As #n Lys Lys Ala Arg Leu          35          #         40          #         45 Arg Ser Ile Phe Pro Gly Gly Gly Asp Lys Th #r Asn Lys Lys Lys Glu      50              #     55              #     60 Lys Glu Arg Pro Glu Ile Ser Leu Pro Ser As #p Phe Glu His Thr Ile  65                  # 70                  # 75                  # 80 His Val Gly Phe Asp Ala Val Thr Gly Glu Ph #e Thr Gly Ile Pro Glu                  85  #                 90  #                 95 Gln Trp Ala Arg Leu Leu Gln Thr Ser Asn Il #e Thr Lys Leu Glu Gln             100       #           105       #           110 Lys Lys Asn Pro Gln Ala Val Leu Asp Val Le #u Lys Phe Tyr Asp Ser         115           #       120           #       125 Lys Glu Thr Val Asn Asn Gln Lys Tyr Met Se #r Phe Thr Ser Gly Asp     130               #   135               #   140 Lys Ser Ala His Gly Tyr Ile Ala Ala His Gl #n Ser Asn Thr Lys Thr 145                 1 #50                 1 #55                 1 #60 Gly Ser Glu Pro Pro Leu Ala Pro Pro Val Se #r Glu Glu Glu Asp Glu                 165   #               170   #               175 Glu Glu Glu Glu Glu Glu Asp Asp Asn Glu Pr #o Pro Pro Val Ile Ala             180       #           185       #           190 Pro Arg Pro Glu His Thr Lys Ser Ile Tyr Th #r Arg Ser Val Val Glu         195           #       200           #       205 Ser Ile Ala Ser Pro Ala Ala Pro Asn Lys Gl #u Asp Ile Pro Pro Ser     210               #   215               #   220 Ala Glu Asn Ala Asn Ser Thr Thr Leu Tyr Ar #g Asn Thr Asp Arg Gln 225                 2 #30                 2 #35                 2 #40 Arg Lys Lys Ser Lys Met Thr Asp Glu Glu Il #e Leu Glu Lys Leu Arg                 245   #               250   #               255 Ser Ile Val Ser Val Gly Asp Pro Lys Lys Ly #s Tyr Thr Arg Leu Glu             260       #           265       #           270 Lys Ile Gly Gln Gly Ala Ser Gly Thr Val Ty #r Thr Ala Leu Asp Ile         275           #       280           #       285 Ala Thr Gly Gln Glu Val Ala Ile Lys Gln Me #t Asn Leu Gln Gln Gln     290               #   295               #   300 Pro Lys Lys Glu Leu Ile Ile Asn Glu Ile Le #u Val Met Arg Glu Asn 305                 3 #10                 3 #15                 3 #20 Lys Asn Pro Asn Ile Val Asn Tyr Leu Asp Se #r Tyr Leu Val Gly Asp                 325   #               330   #               335 Glu Leu Trp Val Val Met Glu Tyr Leu Ala Gl #y Gly Ser Leu Thr Asp             340       #           345       #           350 Val Val Thr Glu Thr Cys Met Asp Val Gly Gl #n Ile Ala Ala Val Cys         355           #       360           #       365 Arg Glu Cys Leu Gln Ala Leu Asp Phe Leu Hi #s Ser Asn Gln Val Ile     370               #   375               #   380 His Arg Asp Ile Lys Ser Asp Asn Ile Leu Le #u Gly Met Asp Gly Ser 385                 3 #90                 3 #95                 4 #00 Val Lys Leu Thr Asp Phe Gly Phe Cys Ala Gl #n Ile Thr Pro Glu Gln                 405   #               410   #               415 Ser Lys Arg Ser Thr Met Val Gly Thr Pro Ty #r Trp Met Ala Pro Glu             420       #           425       #           430 Val Val Thr Arg Lys Ala Tyr Gly Pro Lys Va #l Asp Ile Trp Ser Leu         435           #       440           #       445 Gly Ile Met Ala Ile Glu Met Val Glu Gly Gl #u Pro Pro Tyr Leu Asn     450               #   455               #   460 Glu Asn Pro Leu Arg Ala Leu Tyr Leu Ile Al #a Thr Asn Gly Thr Pro 465                 4 #70                 4 #75                 4 #80 Glu Leu Gln Asn Pro Glu Arg Leu Ser Ala Va #l Phe His Asp Phe Leu                 485   #               490   #               495 Asn Arg Cys Leu Glu Met Asp Val Asp Arg Ar #g Gly Ser Ala Lys Glu             500       #           505       #           510 Leu Leu Gln His Pro Phe Leu Lys Leu Ala Ly #s Pro Leu Ser Ser Leu         515           #       520           #       525 Thr Pro Leu Ile Ile Ala Ala Lys Glu Ala Il #e Lys Asn Ser Ser Arg     530               #   535               #   540 <210> SEQ ID NO 96 <211> LENGTH: 2110 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 96 ggccaagacg gtcggggctg cttgctaact ccaggaacag gtttaagttt tt #gaaactga     60 agtaggtcta cacagtagga actcatgtca tttcttgtaa gtaaaccaga gc #gaatcagg    120 cggtgggtct cggaaaagtt cattgttgag ggcttaagag atttggaact at #ttggagac    180 caatgatgcg agctcagagt caatagcatc cttctctaaa caggaggtca tg #agtagctt    240 tctgccagag ggagggtgtt acgagctgct cactgtgata ggcaaaggat tt #gaggacct    300 gatgactgtg aatctagcaa ggtacaaacc aacaggagag tacgtgactg ta #cggaggat    360 taacctagaa gcttgttcca atgagatggt aacattcttg cagggcgagc tg #catgtctc    420 caaactcttc aaccatccca atatcgtgcc atatcgagcc acttttattg ca #gacaatga    480 gctgtgggtt gtcacatcat tcatggcata cggttctgca aaagatctca tc #tgtacaca    540 cttcatggat ggcatgaatg agctggcgat tgcttacatc ctgcaggggg tg #ctgaaggc    600 cctcgactac atccaccaca tgggatatgt acacaggagt gtcaaagcca gc #cacatcct    660 gatctctgtg gatgggaagg tctacctgtc tggtttgcgc agcaacctca gc #atgataag    720 ccatgggcag cggcagcgag tggtccacga ttttcccaag tacagtgtca ag #gttctgcc    780 gtggctcagc cccgaggtcc tccagcagaa tctccagggt tatgatgcca ag #tctgacat    840 ctacagtgtg ggaatcacag cctgtgaact ggccaacggc catgtcccct tt #aaggatat    900 gcctgccacc cagatgctgc tagagaaact gaacggcaca gtgccctgcc tg #ttggatac    960 cagcaccatc cccgctgagg agctgaccat gagcccttcg cgctcagtgg cc #aactctgg   1020 cctgagtgac agcctgacca ccagcacccc ccggccctcc aacggtgact cg #ccctccca   1080 cccctaccac cgaaccttct ccccccactt ccaccacttt gtggagcagt gc #cttcagcg   1140 caacccggat gccaggccca gtgccagcac cctcctgaac cactctttct tc #aagcagat   1200 caagcgacgt gcctcagagg ctttgcccga attgcttcgt cctgtcaccc cc #atcaccaa   1260 ttttgagggc agccagtctc aggaccacag tggaatcttt ggcctggtaa ca #aacctgga   1320 agagctggag gtggacgatt gggagttctg agcctctgca aactgtgcgc at #tctccagc   1380 cagggatgca gaggccaccc agaggccctt cctgagggcc ggccacattc cc #gccctcct   1440 gggcagattg ggtagaaagg acattcttcc aggaaagttg actgctgact ga #ttgggaaa   1500 gaaaatcctg gagagatact tcactgctcc aaggcttttg agacacaagg ga #atctcaac   1560 aaccagggat caggagggtc caaagccgac attcccagtc ctgtgagctc ag #gtgacctc   1620 ctccgcagaa gagagatgct gctctggccc tgggagctga attccaagcc ca #gggtttgg   1680 ctccttaaac ccgaggaccg ccacctcttc ccagtgcttg cgaccagcct ca #ttctattt   1740 aactttgctc tcagatgcct cagatgctat aggtcagtga aagggcaagt ag #taagctgc   1800 ctgcctccct tccctcagac ctctccctca taattccaga gaagggcatt tc #tgtctttt   1860 taagcacaga ctaaggctgg aacagtccat ccttatccct cttctggctt gg #gccctgac   1920 acctaagtct ttcccacggt ttatgtgtgt gcctcattcc tttcccacca ag #aatccatc   1980 ttagcgcctc ctgccagctg ccctggtgct ttctccaagg gccatcagtg tc #ttgcctag   2040 cttgagggct taagtcctta tgctgtgtta gtttcgttgt cagaacaaat ta #aaattttc   2100 agagacgctg                 #                   #                   #      2110 <210> SEQ ID NO 97 <211> LENGTH: 373 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 97 Met Ser Ser Phe Leu Pro Glu Gly Gly Cys Ty #r Glu Leu Leu Thr Val   1               5  #                 10  #                 15 Ile Gly Lys Gly Phe Glu Asp Leu Met Thr Va #l Asn Leu Ala Arg Tyr              20      #             25      #             30 Lys Pro Thr Gly Glu Tyr Val Thr Val Arg Ar #g Ile Asn Leu Glu Ala          35          #         40          #         45 Cys Ser Asn Glu Met Val Thr Phe Leu Gln Gl #y Glu Leu His Val Ser      50              #     55              #     60 Lys Leu Phe Asn His Pro Asn Ile Val Pro Ty #r Arg Ala Thr Phe Ile  65                  # 70                  # 75                  # 80 Ala Asp Asn Glu Leu Trp Val Val Thr Ser Ph #e Met Ala Tyr Gly Ser                  85  #                 90  #                 95 Ala Lys Asp Leu Ile Cys Thr His Phe Met As #p Gly Met Asn Glu Leu             100       #           105       #           110 Ala Ile Ala Tyr Ile Leu Gln Gly Val Leu Ly #s Ala Leu Asp Tyr Ile         115           #       120           #       125 His His Met Gly Tyr Val His Arg Ser Val Ly #s Ala Ser His Ile Leu     130               #   135               #   140 Ile Ser Val Asp Gly Lys Val Tyr Leu Ser Gl #y Leu Arg Ser Asn Leu 145                 1 #50                 1 #55                 1 #60 Ser Met Ile Ser His Gly Gln Arg Gln Arg Va #l Val His Asp Phe Pro                 165   #               170   #               175 Lys Tyr Ser Val Lys Val Leu Pro Trp Leu Se #r Pro Glu Val Leu Gln             180       #           185       #           190 Gln Asn Leu Gln Gly Tyr Asp Ala Lys Ser As #p Ile Tyr Ser Val Gly         195           #       200           #       205 Ile Thr Ala Cys Glu Leu Ala Asn Gly His Va #l Pro Phe Lys Asp Met     210               #   215               #   220 Pro Ala Thr Gln Met Leu Leu Glu Lys Leu As #n Gly Thr Val Pro Cys 225                 2 #30                 2 #35                 2 #40 Leu Leu Asp Thr Ser Thr Ile Pro Ala Glu Gl #u Leu Thr Met Ser Pro                 245   #               250   #               255 Ser Arg Ser Val Ala Asn Ser Gly Leu Ser As #p Ser Leu Thr Thr Ser             260       #           265       #           270 Thr Pro Arg Pro Ser Asn Gly Asp Ser Pro Se #r His Pro Tyr His Arg         275           #       280           #       285 Thr Phe Ser Pro His Phe His His Phe Val Gl #u Gln Cys Leu Gln Arg     290               #   295               #   300 Asn Pro Asp Ala Arg Pro Ser Ala Ser Thr Le #u Leu Asn His Ser Phe 305                 3 #10                 3 #15                 3 #20 Phe Lys Gln Ile Lys Arg Arg Ala Ser Glu Al #a Leu Pro Glu Leu Leu                 325   #               330   #               335 Arg Pro Val Thr Pro Ile Thr Asn Phe Glu Gl #y Ser Gln Ser Gln Asp             340       #           345       #           350 His Ser Gly Ile Phe Gly Leu Val Thr Asn Le #u Glu Glu Leu Glu Val         355           #       360           #       365 Asp Asp Trp Glu Phe     370 <210> SEQ ID NO 98 <211> LENGTH: 2001 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 98 aaggaagata aaacaaaagc cttctttgga atagatggat ttttgtcact tt #ctgtgtga     60 actaaagtga ttcaatgtct cttttggatt gcttctgcac ttcaagaaca ca #agttgaat    120 cactcagacc tgaaaaacag tctgaaacca gtatccatca atacttggtt ga #tgagccaa    180 ccctttcctg gtcacgtcca tccactagag ccagtgaagt actatgttcc ac #caacgttt    240 ctcactatga gctccaagta gaaataggaa gaggatttga caacttgact tc #tgtccatc    300 ttgcacggca tactcccacg ggaacactgg taactataaa aattacaaat ct #ggaaaact    360 gcaatgaaga acgcctgaaa gctttacaga aagccgtgat tctatcccac tt #tttccggc    420 atcccaatat tacaacttat tggacagttt tcactgttgg cagctggctt tg #ggttattt    480 ctccatttat ggcctatggt tcagcaagtc aactcttgag gacctatttt cc #tgaaggaa    540 tgagtgaaac tttaataaga aacattctct ttggagccgt gagagggttg aa #ctatctgc    600 accaaaatgg ctgtattcac aggagtatta aagccagcca tatcctcatt tc #tggtgatg    660 gcctagtgac cctctctggc ctttcccatc tgcatagttt ggttaagcat gg #acagaggc    720 atagggctgt gtatgatttc ccacagttca gcacatcagt gcagccgtgg ct #gagtccag    780 aactactgag acaggattta catgggtata atgtgaagtc agatatttac ag #tgttggga    840 ttacagcatg tgaattagcc agtgggcagg tgcctttcca ggacatgcat ag #aactcaga    900 tgctgttaca gaaactgaaa ggtcctcctt atagcccatt ggatatcagt at #tttccctc    960 aatcagaatc cagaatgaaa aattcccagt caggtgtaga ctctgggatt gg #agaaagtg   1020 tgcttgtctc cagtggaact cacacagtaa atagtgaccg attacacaca cc #atcctcaa   1080 aaactttctc tcctgccttc tttagcttgg tacagctctg tttgcaacaa ga #tcctgaga   1140 aaaggccatc agcaagcagt ttattgtccc atgttttctt caaacagatg aa #agaagaaa   1200 gccaggattc aatactttca ctgttgcctc ctgcttataa caagccatca at #atcattgc   1260 ctccagtgtt accttggact gagccagaat gtgattttcc tgatgaaaaa ga #ctcatact   1320 gggaattcta gggctgccaa atcattttat gtcctatata cttgacactt tc #tccttgct   1380 gctttttctt ctgtatttct aggtacaaat accagaatta tacttgaaaa ta #cagttggt   1440 gcactggaga atctattatt taaaaccact ctgttcaaag gggcaccagt tt #gtagtccc   1500 tctgtttcgc acagagtact atgacaagga aacatcagaa ttactaatct ag #ctagtgtc   1560 atttattctg gaattttttt ctaagctgtg actaactctt tttatctctc aa #tataattt   1620 ttgagccagt taattttttt cagtattttg ctgtcccttg ggaatgggcc ct #cagaggac   1680 agtgcttcca agtacatctt ctcccagatt ctctggcctt tttaatgagc ta #ttgttaaa   1740 ccaacaggct agtttatctt acatcagacc cttttctggt agagggaaaa tg #tttgtgct   1800 ttcccttttt cttctgttaa tacttatggt aacacctaac tgagcctcac tc #acattaaa   1860 tgattcactt gaaatatata cagaaattgt aatttgcttt tttttaaaaa ag #ggggctaa   1920 agtaacactt tcctacttat gtaaattata gatcctaaat tcacgcaccc cg #tgggagct   1980 caataaagat ttactgaatt g            #                   #                2001 <210> SEQ ID NO 99 <211> LENGTH: 418 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 99 Met Ser Leu Leu Asp Cys Phe Cys Thr Ser Ar #g Thr Gln Val Glu Ser   1               5  #                 10  #                 15 Leu Arg Pro Glu Lys Gln Ser Glu Thr Ser Il #e His Gln Tyr Leu Val              20      #             25      #             30 Asp Glu Pro Thr Leu Ser Trp Ser Arg Pro Se #r Thr Arg Ala Ser Glu          35          #         40          #         45 Val Leu Cys Ser Thr Asn Val Ser His Tyr Gl #u Leu Gln Val Glu Ile      50              #     55              #     60 Gly Arg Gly Phe Asp Asn Leu Thr Ser Val Hi #s Leu Ala Arg His Thr  65                  # 70                  # 75                  # 80 Pro Thr Gly Thr Leu Val Thr Ile Lys Ile Th #r Asn Leu Glu Asn Cys                  85  #                 90  #                 95 Asn Glu Glu Arg Leu Lys Ala Leu Gln Lys Al #a Val Ile Leu Ser His             100       #           105       #           110 Phe Phe Arg His Pro Asn Ile Thr Thr Tyr Tr #p Thr Val Phe Thr Val         115           #       120           #       125 Gly Ser Trp Leu Trp Val Ile Ser Pro Phe Me #t Ala Tyr Gly Ser Ala     130               #   135               #   140 Ser Gln Leu Leu Arg Thr Tyr Phe Pro Glu Gl #y Met Ser Glu Thr Leu 145                 1 #50                 1 #55                 1 #60 Ile Arg Asn Ile Leu Phe Gly Ala Val Arg Gl #y Leu Asn Tyr Leu His                 165   #               170   #               175 Gln Asn Gly Cys Ile His Arg Ser Ile Lys Al #a Ser His Ile Leu Ile             180       #           185       #           190 Ser Gly Asp Gly Leu Val Thr Leu Ser Gly Le #u Ser His Leu His Ser         195           #       200           #       205 Leu Val Lys His Gly Gln Arg His Arg Ala Va #l Tyr Asp Phe Pro Gln     210               #   215               #   220 Phe Ser Thr Ser Val Gln Pro Trp Leu Ser Pr #o Glu Leu Leu Arg Gln 225                 2 #30                 2 #35                 2 #40 Asp Leu His Gly Tyr Asn Val Lys Ser Asp Il #e Tyr Ser Val Gly Ile                 245   #               250   #               255 Thr Ala Cys Glu Leu Ala Ser Gly Gln Val Pr #o Phe Gln Asp Met His             260       #           265       #           270 Arg Thr Gln Met Leu Leu Gln Lys Leu Lys Gl #y Pro Pro Tyr Ser Pro         275           #       280           #       285 Leu Asp Ile Ser Ile Phe Pro Gln Ser Glu Se #r Arg Met Lys Asn Ser     290               #   295               #   300 Gln Ser Gly Val Asp Ser Gly Ile Gly Glu Se #r Val Leu Val Ser Ser 305                 3 #10                 3 #15                 3 #20 Gly Thr His Thr Val Asn Ser Asp Arg Leu Hi #s Thr Pro Ser Ser Lys                 325   #               330   #               335 Thr Phe Ser Pro Ala Phe Phe Ser Leu Val Gl #n Leu Cys Leu Gln Gln             340       #           345       #           350 Asp Pro Glu Lys Arg Pro Ser Ala Ser Ser Le #u Leu Ser His Val Phe         355           #       360           #       365 Phe Lys Gln Met Lys Glu Glu Ser Gln Asp Se #r Ile Leu Ser Leu Leu     370               #   375               #   380 Pro Pro Ala Tyr Asn Lys Pro Ser Ile Ser Le #u Pro Pro Val Leu Pro 385                 3 #90                 3 #95                 4 #00 Trp Thr Glu Pro Glu Cys Asp Phe Pro Asp Gl #u Lys Asp Ser Tyr Trp                 405   #               410   #               415 Glu Phe <210> SEQ ID NO 100 <211> LENGTH: 311 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 100 tcaacaggga cgattacgag ctgcaggagg tgatcgggag tggagcaact gc #tgtagtcc     60 aagcagctta ttgtgcccct aaaaaggaga aagtggcaat caaacggata aa #ccttgaga    120 aatgtcaaac tagcatggat gaactcctga aagaaattca agccatgagt ca #atgccatc    180 atcctaatat tgtatcttac tacacatctt ttgtggtaaa agatgagctg tg #gcttgtca    240 tgaagctgct aagtggaggt tctgttctgg atattattaa gcacattgtg gc #aaaagggg    300 aacacaaaag t                #                   #                   #      311 <210> SEQ ID NO 101 <211> LENGTH: 103 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 101 Asn Arg Asp Asp Tyr Glu Leu Gln Glu Val Il #e Gly Ser Gly Ala Thr   1               5  #                 10  #                 15 Ala Val Val Gln Ala Ala Tyr Cys Ala Pro Ly #s Lys Glu Lys Val Ala              20      #             25      #             30 Ile Lys Arg Ile Asn Leu Glu Lys Cys Gln Th #r Ser Met Asp Glu Leu          35          #         40          #         45 Leu Lys Glu Ile Gln Ala Met Ser Gln Cys Hi #s His Pro Asn Ile Val      50              #     55              #     60 Ser Tyr Tyr Thr Ser Phe Val Val Lys Asp Gl #u Leu Trp Leu Val Met  65                  # 70                  # 75                  # 80 Lys Leu Leu Ser Gly Gly Ser Val Leu Asp Il #e Ile Lys His Ile Val                  85  #                 90  #                 95 Ala Lys Gly Glu His Lys Ser             100 <210> SEQ ID NO 102 <211> LENGTH: 2806 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 102 cgggagtgtc cgcggtggtg gcggtgcaag agagctgaag gaggcgcgag gg #cgcggagt     60 tccaggccga gcagttaggc cgcgagcgac tgcggcgccg agccgatgag ta #acccgaag    120 cccctagagg agtggtcacc tgcctgaggg cacttctgtc ccaccagcat ca #gaccaggc    180 cgcaccgagt ccccggcacc atgtttggga agaggaagaa gcgggtggag at #ctccgcgc    240 cgtccaactt cgagcaccgc gtgcacacgg gcttcgacca gcacgagcag aa #gttcacgg    300 ggctgccccg ccagtggcag agcctgatcg aggagtcggc tcgccggccc aa #gcccctcg    360 tcgaccccgc ctgcatcacc tccatccagc ccggggcccc caagaccatc gt #gcggggca    420 gcaaaggtgc caaagatggg gccctcacgc tgctgctgga cgagtttgag aa #catgtcgg    480 tgacacgctc caactccctg cggagagaca gcccgccgcc gcccgcccgt gc #ccgccagg    540 aaaatgggat gccagaggag ccggccacca cggccagagg gggcccaggg aa #ggcaggca    600 gccgaggccg gttcgccggt cacagcgagg cgggtggcgg cagtggtgac ag #gcgacggg    660 cggggccaga gaagaggccc aagtcttcca gggagggctc agggggtccc ca #ggagtcct    720 cccgggacaa acgccccctc tccgggcctg atgtcggcac cccccagcct gc #tggtctgg    780 ccagtggggc gaaactggca gctggccggc cctttaacac ctacccgagg gc #tgacacgg    840 accacccatc ccggggtgcc cagggggagc ctcatgacgt ggcccctaac gg #gccatcag    900 cggggggcct ggccatcccc cagtcctcct cctcctcctc ccggcctccc ac #ccgagccc    960 gaggtgcccc cagccctgga gtgctgggac cccacgcctc agagccccag ct #ggcccctc   1020 cagcctgcac ccccgccgcc cctgctgttc ctgggccccc tggcccccgc tc #accacagc   1080 gggagccaca gcgagtatcc catgagcagt tccgggctgc cctgcagctg gt #ggtggacc   1140 caggcgaccc ccgctcctac ctggacaact tcatcaagat tggcgagggc tc #cacgggca   1200 tcgtgtgcat cgccaccgtg cgcagctcgg gcaagctggt ggccgtcaag aa #gatggacc   1260 tgcgcaagca gcagaggcgc gagctgctct tcaacgaggt ggtaatcatg ag #ggactacc   1320 agcacgagaa tgtggtggag atgtacaaca gctacctggt gggggacgag ct #ctgggtgg   1380 tcatggagtt cctggaagga ggcgccctca ccgacatcgt cacccacacc ag #gatgaacg   1440 aggagcagat cgcggccgtg tgccttgcag tgctgcaggc cctgtcggtg ct #ccacgccc   1500 agggcgtcat ccaccgggac atcaagagcg actcgatcct gctgacccat ga #tggcaggg   1560 tgaagctgtc agactttggg ttctgcgccc aggtgagcaa ggaagtgccc cg #aaggaagt   1620 cgctggtcgg cacgccctac tggatggccc cagagctcat ctcccgcctt cc #ctacgggc   1680 cagaggtaga catctggtcg ctggggataa tggtgattga gatggtggac gg #agagcccc   1740 cctacttcaa cgagccaccc ctcaaagcca tgaagatgat tcgggacaac ct #gccacccc   1800 gactgaagaa cctgcacaag gtgtcgccat ccctgaaggg cttcctggac cg #cctgctgg   1860 tgcgagaccc tgcccagcgg gccacggcag ccgagctgct gaagcaccca tt #cctggcca   1920 aggcagggcc gcctgccagc atcgtgcccc tcatgcgcca gaaccgcacc ag #atgaggcc   1980 cagcgccctt cccctcaacc aaagagcccc cccgggtcac ccccgcccca ct #gaggccag   2040 tagggggcca ggcctcccac tcctcccagc ccgggagatg ctccgcgtgg ca #ccaccctc   2100 cttgctgggg gtagatgaga ccctactact gaactccagt tttgatctcg tg #acttttag   2160 aaaaacacag ggactcgtgg gagcaagcga ggctcccagg acccccaccc tc #tgggacag   2220 gccctccccc atgttcttct gtctccagga agggcagcgg ccctcccatc ac #tggaagtc   2280 tgcagtgggg gtcgctgggg gtggagagaa cactaagagg tgaacatgta tg #agtgtgtg   2340 cacgcgtgtg agtgtgcatg tgtgtgtgtg tgcaaaggtc cagccacccc gt #cctccagc   2400 ccgcaagggg tgtctggcgc cttgcctgac acccagcccc ctctccccct ga #gccattgt   2460 gggggtcgat catgaatgtc cgaagagtgg ccttttcccg tagccctgcg cc #ccctttct   2520 gtggctggat ggggagacag gtcagggccc cccaccctct ccagcccctg ca #gcaaatga   2580 ctactgcacc tggacagcct cctcttttct agaagtctat ttatattgtc at #tttataac   2640 actctagccc ctgcccttat tgggggacag atggtccctg tcctgcgggg tg #gccctggc   2700 agaaccactg cctgaagaac caggttcctg cccggtcagc gcagccccag cc #cgcccacc   2760 cctgcctcga gttagtttta caattaaaac attgtcttgt tttgtg    #               2806 <210> SEQ ID NO 103 <211> LENGTH: 591 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 103 Met Phe Gly Lys Arg Lys Lys Arg Val Glu Il #e Ser Ala Pro Ser Asn   1               5  #                 10  #                 15 Phe Glu His Arg Val His Thr Gly Phe Asp Gl #n His Glu Gln Lys Phe              20      #             25      #             30 Thr Gly Leu Pro Arg Gln Trp Gln Ser Leu Il #e Glu Glu Ser Ala Arg          35          #         40          #         45 Arg Pro Lys Pro Leu Val Asp Pro Ala Cys Il #e Thr Ser Ile Gln Pro      50              #     55              #     60 Gly Ala Pro Lys Thr Ile Val Arg Gly Ser Ly #s Gly Ala Lys Asp Gly  65                  # 70                  # 75                  # 80 Ala Leu Thr Leu Leu Leu Asp Glu Phe Glu As #n Met Ser Val Thr Arg                  85  #                 90  #                 95 Ser Asn Ser Leu Arg Arg Asp Ser Pro Pro Pr #o Pro Ala Arg Ala Arg             100       #           105       #           110 Gln Glu Asn Gly Met Pro Glu Glu Pro Ala Th #r Thr Ala Arg Gly Gly         115           #       120           #       125 Pro Gly Lys Ala Gly Ser Arg Gly Arg Phe Al #a Gly His Ser Glu Ala     130               #   135               #   140 Gly Gly Gly Ser Gly Asp Arg Arg Arg Ala Gl #y Pro Glu Lys Arg Pro 145                 1 #50                 1 #55                 1 #60 Lys Ser Ser Arg Glu Gly Ser Gly Gly Pro Gl #n Glu Ser Ser Arg Asp                 165   #               170   #               175 Lys Arg Pro Leu Ser Gly Pro Asp Val Gly Th #r Pro Gln Pro Ala Gly             180       #           185       #           190 Leu Ala Ser Gly Ala Lys Leu Ala Ala Gly Ar #g Pro Phe Asn Thr Tyr         195           #       200           #       205 Pro Arg Ala Asp Thr Asp His Pro Ser Arg Gl #y Ala Gln Gly Glu Pro     210               #   215               #   220 His Asp Val Ala Pro Asn Gly Pro Ser Ala Gl #y Gly Leu Ala Ile Pro 225                 2 #30                 2 #35                 2 #40 Gln Ser Ser Ser Ser Ser Ser Arg Pro Pro Th #r Arg Ala Arg Gly Ala                 245   #               250   #               255 Pro Ser Pro Gly Val Leu Gly Pro His Ala Se #r Glu Pro Gln Leu Ala             260       #           265       #           270 Pro Pro Ala Cys Thr Pro Ala Ala Pro Ala Va #l Pro Gly Pro Pro Gly         275           #       280           #       285 Pro Arg Ser Pro Gln Arg Glu Pro Gln Arg Va #l Ser His Glu Gln Phe     290               #   295               #   300 Arg Ala Ala Leu Gln Leu Val Val Asp Pro Gl #y Asp Pro Arg Ser Tyr 305                 3 #10                 3 #15                 3 #20 Leu Asp Asn Phe Ile Lys Ile Gly Glu Gly Se #r Thr Gly Ile Val Cys                 325   #               330   #               335 Ile Ala Thr Val Arg Ser Ser Gly Lys Leu Va #l Ala Val Lys Lys Met             340       #           345       #           350 Asp Leu Arg Lys Gln Gln Arg Arg Glu Leu Le #u Phe Asn Glu Val Val         355           #       360           #       365 Ile Met Arg Asp Tyr Gln His Glu Asn Val Va #l Glu Met Tyr Asn Ser     370               #   375               #   380 Tyr Leu Val Gly Asp Glu Leu Trp Val Val Me #t Glu Phe Leu Glu Gly 385                 3 #90                 3 #95                 4 #00 Gly Ala Leu Thr Asp Ile Val Thr His Thr Ar #g Met Asn Glu Glu Gln                 405   #               410   #               415 Ile Ala Ala Val Cys Leu Ala Val Leu Gln Al #a Leu Ser Val Leu His             420       #           425       #           430 Ala Gln Gly Val Ile His Arg Asp Ile Lys Se #r Asp Ser Ile Leu Leu         435           #       440           #       445 Thr His Asp Gly Arg Val Lys Leu Ser Asp Ph #e Gly Phe Cys Ala Gln     450               #   455               #   460 Val Ser Lys Glu Val Pro Arg Arg Lys Ser Le #u Val Gly Thr Pro Tyr 465                 4 #70                 4 #75                 4 #80 Trp Met Ala Pro Glu Leu Ile Ser Arg Leu Pr #o Tyr Gly Pro Glu Val                 485   #               490   #               495 Asp Ile Trp Ser Leu Gly Ile Met Val Ile Gl #u Met Val Asp Gly Glu             500       #           505       #           510 Pro Pro Tyr Phe Asn Glu Pro Pro Leu Lys Al #a Met Lys Met Ile Arg         515           #       520           #       525 Asp Asn Leu Pro Pro Arg Leu Lys Asn Leu Hi #s Lys Val Ser Pro Ser     530               #   535               #   540 Leu Lys Gly Phe Leu Asp Arg Leu Leu Val Ar #g Asp Pro Ala Gln Arg 545                 5 #50                 5 #55                 5 #60 Ala Thr Ala Ala Glu Leu Leu Lys His Pro Ph #e Leu Ala Lys Ala Gly                 565   #               570   #               575 Pro Pro Ala Ser Ile Val Pro Leu Met Arg Gl #n Asn Arg Thr Arg             580       #           585       #           590 <210> SEQ ID NO 104 <211> LENGTH: 3684 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 104 atggcgggac ctgggggctg gagggacagg gaggtcacgg atctgggcca cc #tgccggat     60 ccaactggaa tattctcact agataaaacc attggccttg gtacttatgg ca #gaatctat    120 ttgggacttc atgagaagac tggtgcattt acagctgtta aagtgatgaa cg #ctcgtaag    180 gatgaggaag aggatctcag gactgaactc aaccttctga ggaagtactc tt #tccacaaa    240 aacattgtgt ccttctatgg agcatttttc aagctgagtc cccctggtca gc #ggcaccaa    300 ctttggatgg tgatggagtt atgtgcagca ggttcggtca ctgatgtagt ga #gaatgacc    360 agtaatcaga gtttaaaaga agattggatt gcttatatct gccgagaaat cc #ttcagggc    420 ttagctcacc ttcacgcaca ccgagtaatt caccgggaca tcaaaggtca ga #atgtgctg    480 ctgactcata atgctgaagt aaaactggtt gattttggag tgagtgccca gg #tgagcaga    540 actaatggaa gaaggaatag tttcattggg acaccatact ggatggcacc tg #aggtgatt    600 gactgtgatg aggacccaag acgctcctat gattacagaa gtgatgtgtg gt #ctgtggga    660 attactgcca ttgaaatggc tgaaggagcc cctcctctgt gtaaccttca ac #ccttggaa    720 gctctcttcg ttattttgcg ggaatctgct cccacagtca aatccagcgg at #ggtcccgt    780 aagttccaca atttcatgga aaagtgtacg ataaaaaatt tcctgtttcg tc #ctacttct    840 gcaaacatgc ttcaacaccc atttgttcgg gatataaaaa atgaacgaca tg #ttgttgag    900 tcattaacaa ggcatcttac tggaatcatt aaaaaaagac agaaaaaaga ac #aggcacgg    960 gagaaaaaat caaaagtttc tactctgagg caagcactgg caaaaagact at #caccaaag   1020 aggttcaggg caaagtcatc atggagacct gaaaagcttg aactctcgga tt #tagaagcc   1080 cgcaggcaaa ggcgccaacg cagatgggaa gatatcttta atcagcatga gg #aagaattg   1140 agacaagttg ataaagacaa agaagatgaa tcatcagaca atgatgaagt at #ttcattcg   1200 attcaggctg aagtccagat agagccattg aagccataca tttcaaatcc ta #aaaaaatt   1260 gaggttcaag agagatctcc ttctgtgcct aacaaccagg atcatgcaca tc #atgtcaag   1320 ttctcttcaa gcgttcctca gcggtctctt ttggaacaag ctcagaagcc ca #ttgacatc   1380 agacaaagga gttcgcaaaa tcgtcaaaat tggctggcag catcaggtga tt #caaagcac   1440 aaaattttag caggcaaaac acagagctac tgtttaacaa tttatatttc ag #aagtcaag   1500 aaagaagaat ttcaagaagg aatgaatcaa aagtgtcagg gagcccaagt ag #gattagga   1560 cctgaaggcc attgtatttg gcaattgggt gaatcttctt ctgaggaaga aa #gtcctgtg   1620 actggaagga ggtctcagtc atcaccacct tattctacta ttgatcagaa gt #tgctggtt   1680 gacatccatg ttccagatgg atttaaagta ggaaaaatat caccccctgt at #acttgaca   1740 aacgaatggg taggctataa tgcactctct gaaatcttcc ggaatgattg gt #taactccg   1800 gcacctgtca ttcagccacc tgaagaggat ggtgattatg ttgaactcta tg #atgccagt   1860 gctgatactg atggtgatga tgatgatgag tctaatgata cttttgaaga ta #cctatgat   1920 catgccaatg gcaatgatga cttggataac caggttgatc aggctaatga tg #tttgtaaa   1980 gaccatgatg atgacaacaa taagtttgtt gatgatgtaa ataataatta tt #atgaggcg   2040 cctagttgtc caagggcaag ctatggcaga gatggaagct gcaagcaaga tg #gttatgat   2100 ggaagtcgtg gaaaagagga agcctacaga ggctatggaa gccatacagc ca #atagaagc   2160 catggaggaa gtgcagccag tgaggacaat gcagccattg gagatcagga ag #aacatgca   2220 gccaatatag gcagtgaaag aagaggcagt gagggtgatg gaggtaaggg ag #tcgttcga   2280 accagtgaag agagtggagc ccttggactc aatggagaag aaaattgctc ag #agacagat   2340 ggtccaggat tgaagagacc tgcgtctcag gactttgaat atctacagga gg #agccaggt   2400 ggtggaaatg aggcctcaaa tgccattgac tcaggtgctg caccgtcagc ac #ctgatcat   2460 gagagtgaca ataaggacat atcagaatca tcaacacaat cagatttttc tg #ccaatcac   2520 tcatctcctt ccaaaggttc tgggatgtct gctgatgcta actttgccag tg #ccatctta   2580 tacgctggat tcgtagaagt acctgaggaa tcacctaagc aaccctctga ag #tcaatgtt   2640 aacccactct atgtctctcc tgcatgtaaa aaaccactaa tccacatgta tg #aaaaggag   2700 ttcacttctg agatctgctg tggttctttg tggggagtca atttgctgtt gg #gaacccga   2760 tctaatctat atctgatgga cagaagtgga aaggctgaca ttactaaact ta #taaggcga   2820 agaccattcc gccagattca agtcttagag ccactcaatt tgctgattac ca #tctcaggt   2880 cataagaaca gacttcgggt gtatcatctg acctggttga ggaacaagat tt #tgaataat   2940 gatccagaaa gtaaaagaag gcaagaagaa atgctgaaga cagaggaagc ct #gcaaagct   3000 attgataagt taacaggctg tgaacacttc agtgtcctcc aacatgaaga aa #caacatat   3060 attgcaattg ctttgaaatc atcaattcac ctttatgcat gggcaccaaa gt #cctttgat   3120 gaaagcactg ctattaaagt atttccaaca cttgatcata agccagtgac ag #ttgacctg   3180 gctattggtt ctgaaaaaag actaaagatt ttcttcagct cagcagatgg at #atcacctc   3240 atcgatgcag aatctgaggt tatgtctgat gtgaccctgc caaagaatcc cc #tggaaatc   3300 attataccac agaatatcat cattttacct gattgcttgg gaattggcat ga #tgctcacc   3360 ttcaatgctg aagccctctc tgtggaagca aatgaacaac tcttcaagaa ga #tccttgaa   3420 atgtggaaag acataccatc ttctatagct tttgaatgta cacagcgaac ca #caggatgg   3480 ggccaaaagg ccattgaagt gcgctctttg caatccaggg ttctggaaag tg #agctgaag   3540 cgcaggtcaa ttaagaagct gagattcctg tgcacccggg gtgacaagct gt #tctttacc   3600 tctaccctgc gcaatcacca cagccgggtt tacttcatga cacttggaaa ac #ttgaagag   3660 ctccaaagca attatgatgt ctaa           #                   #              3684 <210> SEQ ID NO 105 <211> LENGTH: 1227 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 105 Met Ala Gly Pro Gly Gly Trp Arg Asp Arg Gl #u Val Thr Asp Leu Gly   1               5  #                 10  #                 15 His Leu Pro Asp Pro Thr Gly Ile Phe Ser Le #u Asp Lys Thr Ile Gly              20      #             25      #             30 Leu Gly Thr Tyr Gly Arg Ile Tyr Leu Gly Le #u His Glu Lys Thr Gly          35          #         40          #         45 Ala Phe Thr Ala Val Lys Val Met Asn Ala Ar #g Lys Asp Glu Glu Glu      50              #     55              #     60 Asp Leu Arg Thr Glu Leu Asn Leu Leu Arg Ly #s Tyr Ser Phe His Lys  65                  # 70                  # 75                  # 80 Asn Ile Val Ser Phe Tyr Gly Ala Phe Phe Ly #s Leu Ser Pro Pro Gly                  85  #                 90  #                 95 Gln Arg His Gln Leu Trp Met Val Met Glu Le #u Cys Ala Ala Gly Ser             100       #           105       #           110 Val Thr Asp Val Val Arg Met Thr Ser Asn Gl #n Ser Leu Lys Glu Asp         115           #       120           #       125 Trp Ile Ala Tyr Ile Cys Arg Glu Ile Leu Gl #n Gly Leu Ala His Leu     130               #   135               #   140 His Ala His Arg Val Ile His Arg Asp Ile Ly #s Gly Gln Asn Val Leu 145                 1 #50                 1 #55                 1 #60 Leu Thr His Asn Ala Glu Val Lys Leu Val As #p Phe Gly Val Ser Ala                 165   #               170   #               175 Gln Val Ser Arg Thr Asn Gly Arg Arg Asn Se #r Phe Ile Gly Thr Pro             180       #           185       #           190 Tyr Trp Met Ala Pro Glu Val Ile Asp Cys As #p Glu Asp Pro Arg Arg         195           #       200           #       205 Ser Tyr Asp Tyr Arg Ser Asp Val Trp Ser Va #l Gly Ile Thr Ala Ile     210               #   215               #   220 Glu Met Ala Glu Gly Ala Pro Pro Leu Cys As #n Leu Gln Pro Leu Glu 225                 2 #30                 2 #35                 2 #40 Ala Leu Phe Val Ile Leu Arg Glu Ser Ala Pr #o Thr Val Lys Ser Ser                 245   #               250   #               255 Gly Trp Ser Arg Lys Phe His Asn Phe Met Gl #u Lys Cys Thr Ile Lys             260       #           265       #           270 Asn Phe Leu Phe Arg Pro Thr Ser Ala Asn Me #t Leu Gln His Pro Phe         275           #       280           #       285 Val Arg Asp Ile Lys Asn Glu Arg His Val Va #l Glu Ser Leu Thr Arg     290               #   295               #   300 His Leu Thr Gly Ile Ile Lys Lys Arg Gln Ly #s Lys Glu Gln Ala Arg 305                 3 #10                 3 #15                 3 #20 Glu Lys Lys Ser Lys Val Ser Thr Leu Arg Gl #n Ala Leu Ala Lys Arg                 325   #               330   #               335 Leu Ser Pro Lys Arg Phe Arg Ala Lys Ser Se #r Trp Arg Pro Glu Lys             340       #           345       #           350 Leu Glu Leu Ser Asp Leu Glu Ala Arg Arg Gl #n Arg Arg Gln Arg Arg         355           #       360           #       365 Trp Glu Asp Ile Phe Asn Gln His Glu Glu Gl #u Leu Arg Gln Val Asp     370               #   375               #   380 Lys Asp Lys Glu Asp Glu Ser Ser Asp Asn As #p Glu Val Phe His Ser 385                 3 #90                 3 #95                 4 #00 Ile Gln Ala Glu Val Gln Ile Glu Pro Leu Ly #s Pro Tyr Ile Ser Asn                 405   #               410   #               415 Pro Lys Lys Ile Glu Val Gln Glu Arg Ser Pr #o Ser Val Pro Asn Asn             420       #           425       #           430 Gln Asp His Ala His His Val Lys Phe Ser Se #r Ser Val Pro Gln Arg         435           #       440           #       445 Ser Leu Leu Glu Gln Ala Gln Lys Pro Ile As #p Ile Arg Gln Arg Ser     450               #   455               #   460 Ser Gln Asn Arg Gln Asn Trp Leu Ala Ala Se #r Gly Asp Ser Lys His 465                 4 #70                 4 #75                 4 #80 Lys Ile Leu Ala Gly Lys Thr Gln Ser Tyr Cy #s Leu Thr Ile Tyr Ile                 485   #               490   #               495 Ser Glu Val Lys Lys Glu Glu Phe Gln Glu Gl #y Met Asn Gln Lys Cys             500       #           505       #           510 Gln Gly Ala Gln Val Gly Leu Gly Pro Glu Gl #y His Cys Ile Trp Gln         515           #       520           #       525 Leu Gly Glu Ser Ser Ser Glu Glu Glu Ser Pr #o Val Thr Gly Arg Arg     530               #   535               #   540 Ser Gln Ser Ser Pro Pro Tyr Ser Thr Ile As #p Gln Lys Leu Leu Val 545                 5 #50                 5 #55                 5 #60 Asp Ile His Val Pro Asp Gly Phe Lys Val Gl #y Lys Ile Ser Pro Pro                 565   #               570   #               575 Val Tyr Leu Thr Asn Glu Trp Val Gly Tyr As #n Ala Leu Ser Glu Ile             580       #           585       #           590 Phe Arg Asn Asp Trp Leu Thr Pro Ala Pro Va #l Ile Gln Pro Pro Glu         595           #       600           #       605 Glu Asp Gly Asp Tyr Val Glu Leu Tyr Asp Al #a Ser Ala Asp Thr Asp     610               #   615               #   620 Gly Asp Asp Asp Asp Glu Ser Asn Asp Thr Ph #e Glu Asp Thr Tyr Asp 625                 6 #30                 6 #35                 6 #40 His Ala Asn Gly Asn Asp Asp Leu Asp Asn Gl #n Val Asp Gln Ala Asn                 645   #               650   #               655 Asp Val Cys Lys Asp His Asp Asp Asp Asn As #n Lys Phe Val Asp Asp             660       #           665       #           670 Val Asn Asn Asn Tyr Tyr Glu Ala Pro Ser Cy #s Pro Arg Ala Ser Tyr         675           #       680           #       685 Gly Arg Asp Gly Ser Cys Lys Gln Asp Gly Ty #r Asp Gly Ser Arg Gly     690               #   695               #   700 Lys Glu Glu Ala Tyr Arg Gly Tyr Gly Ser Hi #s Thr Ala Asn Arg Ser 705                 7 #10                 7 #15                 7 #20 His Gly Gly Ser Ala Ala Ser Glu Asp Asn Al #a Ala Ile Gly Asp Gln                 725   #               730   #               735 Glu Glu His Ala Ala Asn Ile Gly Ser Glu Ar #g Arg Gly Ser Glu Gly             740       #           745       #           750 Asp Gly Gly Lys Gly Val Val Arg Thr Ser Gl #u Glu Ser Gly Ala Leu         755           #       760           #       765 Gly Leu Asn Gly Glu Glu Asn Cys Ser Glu Th #r Asp Gly Pro Gly Leu     770               #   775               #   780 Lys Arg Pro Ala Ser Gln Asp Phe Glu Tyr Le #u Gln Glu Glu Pro Gly 785                 7 #90                 7 #95                 8 #00 Gly Gly Asn Glu Ala Ser Asn Ala Ile Asp Se #r Gly Ala Ala Pro Ser                 805   #               810   #               815 Ala Pro Asp His Glu Ser Asp Asn Lys Asp Il #e Ser Glu Ser Ser Thr             820       #           825       #           830 Gln Ser Asp Phe Ser Ala Asn His Ser Ser Pr #o Ser Lys Gly Ser Gly         835           #       840           #       845 Met Ser Ala Asp Ala Asn Phe Ala Ser Ala Il #e Leu Tyr Ala Gly Phe     850               #   855               #   860 Val Glu Val Pro Glu Glu Ser Pro Lys Gln Pr #o Ser Glu Val Asn Val 865                 8 #70                 8 #75                 8 #80 Asn Pro Leu Tyr Val Ser Pro Ala Cys Lys Ly #s Pro Leu Ile His Met                 885   #               890   #               895 Tyr Glu Lys Glu Phe Thr Ser Glu Ile Cys Cy #s Gly Ser Leu Trp Gly             900       #           905       #           910 Val Asn Leu Leu Leu Gly Thr Arg Ser Asn Le #u Tyr Leu Met Asp Arg         915           #       920           #       925 Ser Gly Lys Ala Asp Ile Thr Lys Leu Ile Ar #g Arg Arg Pro Phe Arg     930               #   935               #   940 Gln Ile Gln Val Leu Glu Pro Leu Asn Leu Le #u Ile Thr Ile Ser Gly 945                 9 #50                 9 #55                 9 #60 His Lys Asn Arg Leu Arg Val Tyr His Leu Th #r Trp Leu Arg Asn Lys                 965   #               970   #               975 Ile Leu Asn Asn Asp Pro Glu Ser Lys Arg Ar #g Gln Glu Glu Met Leu             980       #           985       #           990 Lys Thr Glu Glu Ala Cys Lys Ala Ile Asp Ly #s Leu Thr Gly Cys Glu         995           #      1000            #     1005 His Phe Ser Val Leu Gln His Glu Glu Thr Th #r Tyr Ile Ala Ile Ala    1010               #  1015                # 1020 Leu Lys Ser Ser Ile His Leu Tyr Ala Trp Al #a Pro Lys Ser Phe Asp 1025               1030  #               1035   #              1040 Glu Ser Thr Ala Ile Lys Val Phe Pro Thr Le #u Asp His Lys Pro Val                1045   #              1050    #             1055 Thr Val Asp Leu Ala Ile Gly Ser Glu Lys Ar #g Leu Lys Ile Phe Phe            1060       #          1065        #         1070 Ser Ser Ala Asp Gly Tyr His Leu Ile Asp Al #a Glu Ser Glu Val Met        1075           #      1080            #     1085 Ser Asp Val Thr Leu Pro Lys Asn Pro Leu Gl #u Ile Ile Ile Pro Gln    1090               #  1095                # 1100 Asn Ile Ile Ile Leu Pro Asp Cys Leu Gly Il #e Gly Met Met Leu Thr 1105               1110  #               1115   #              1120 Phe Asn Ala Glu Ala Leu Ser Val Glu Ala As #n Glu Gln Leu Phe Lys                1125   #              1130    #             1135 Lys Ile Leu Glu Met Trp Lys Asp Ile Pro Se #r Ser Ile Ala Phe Glu            1140       #          1145        #         1150 Cys Thr Gln Arg Thr Thr Gly Trp Gly Gln Ly #s Ala Ile Glu Val Arg        1155           #      1160            #     1165 Ser Leu Gln Ser Arg Val Leu Glu Ser Glu Le #u Lys Arg Arg Ser Ile    1170               #  1175                # 1180 Lys Lys Leu Arg Phe Leu Cys Thr Arg Gly As #p Lys Leu Phe Phe Thr 1185               1190  #               1195   #              1200 Ser Thr Leu Arg Asn His His Ser Arg Val Ty #r Phe Met Thr Leu Gly                1205   #              1210    #             1215 Lys Leu Glu Glu Leu Gln Ser Asn Tyr Asp Va #l            1220       #          1225 <210> SEQ ID NO 106 <211> LENGTH: 2962 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 106 cgaagccaca gcccgagccc gagcccgagc ccgagccggc gccaccgcgc cc #ccggccat     60 ggcttttgcc aatttccgcc gcatcctgcg cctgtctacc ttcgagaaga ga #aagtcccg    120 cgaatatgag cacgtccgcc gcgacctgga ccccaacgag gtgtgggaga tc #gtgggcga    180 gctgggcgac ggcgccttcg gcaaggttta caaggccaag aataaggaga cg #ggtgcttt    240 ggctgcggcc aaagtcattg aaaccaagag tgaggaggag ctggaggact ac #atcgtgga    300 gattgagatc ctggccacct gcgaccaccc ctacattgtg aagctcctgg ga #gcctacta    360 tcacgacggg aagctgtgga tcatgattga gttctgtcca gggggagccg tg #gacgccat    420 catgctggag ctggacagag gcctcacgga gccccagata caggtggttt gc #cgccagat    480 gctagaagcc ctcaacttcc tgcacagcaa gaggatcatc caccgagatc tg #aaagctgg    540 caacgtgctg atgaccctcg agggagacat caggctggct gactttggtg tg #tctgccaa    600 gaatctgaag actctacaga aacgagattc cttcatcggc acgccttact gg #atggcccc    660 cgaggtggtc atgtgtgaga ccatgaaaga cacgccctac gactacaaag cc #gacatctg    720 gtccctgggc atcacgctga ttgagatggc ccagatcgag ccgccacacc ac #gagctcaa    780 ccccatgcgg gtcctgctaa agatcgccaa gtcggaccct cccacgctgc tc #acgccctc    840 caagtggtct gtagagttcc gtgacttcct gaagatagcc ctggataaga ac #ccagaaac    900 ccgacccagt gccgcgcagc tgctggagca tcccttcgtc agcagcatca cc #agtaacaa    960 ggctctgcgg gagctggtgg ctgaggccaa ggccgaggtg atggaagaga tc #gaagacgg   1020 ccgggatgag ggggaagagg aggacgccgt ggatgccgcc tccaccctgg ag #aaccatac   1080 tcagaactcc tctgaggtga gtccgccaag cctcaatgct gacaagcctc tc #gaggagtc   1140 accttccacc ccgctggcac ccagccagtc tcaggacagt gtgaatgagc cc #tgcagcca   1200 gccctctggg gacagatccc tccaaaccac cagtccccca gtcgtggccc ct #ggaaatga   1260 gaacggcctg gcagtgcctg tgcccctgcg gaagtcccga cccgtgtcaa tg #gatgccag   1320 aattcaggta gcccaggaga agcaagttgc tgagcagggt ggggacctca gc #ccagcagc   1380 caacagatct caaaaggcca gccagagccg gcccaacagc agcgccctgg ag #accttggg   1440 tggggagaag ctggccaatg gcagcctgga gccacctgcc caggcagctc ca #gggccttc   1500 caagagggac tcggactgca gcagcctctg cacctctgag agcatggact at #ggtaccaa   1560 tctctccact gacctgtcgc tgaacaaaga gatgggctct ctgtccatca ag #gacccgaa   1620 actgtacaaa aaaaccctca agcggacacg caaatttgtg gtggatggtg tg #gaggtgag   1680 catcaccacc tccaagatca tcagcgaaga tgagaagaag gatgaggaga tg #agatttct   1740 caggcgccag gaactccgag agcttcggct gctccagaaa gaagagcatc gg #aaccagac   1800 ccagctgagt aacaagcatg agctgcagct ggagcaaatg cataaacgtt tt #gaacagga   1860 aatcaacgcc aagaagaagt tctttgacac ggaattagag aacctggagc gt #cagcaaaa   1920 gcagcaagtg gagaagatgg agcaagacca tgccgtgcgc cgccgggagg ag #gccaggcg   1980 gatccgcctg gagcaggatc gggactacac caggttccaa gagcagctca aa #ctgatgaa   2040 gaaagaggtg aagaacgagg tggagaagct cccccgacag cagcggaagg aa #agcatgaa   2100 gcagaagatg gaggagcaca cgcagaaaaa gcagcttctt gaccgggact tt #gtagccaa   2160 gcagaaggag gacctggagc tggccatgaa gaggctcacc accgacaaca gg #cgggagat   2220 ctgtgacaag gagcgcgagt gcctcatgaa gaagcaggag ctccttcgag ac #cgggaagc   2280 agccctgtgg gagatggaag agcaccagct gcaggagagg caccagctgg tg #aagcagca   2340 gctcaaagac cagtacttcc tccagcggca cgagctgctg cgcaagcatg ag #aaggagcg   2400 ggagcagatg cagcgctaca accagcgcat gatagagcag ctgaaggtgc gg #cagcaaca   2460 ggaaaaggcg cggctgccca agatccagag gagtgagggc aagacgcgca tg #gccatgta   2520 caagaagagc ctccacatca acggcggggg cagcgcagct gagcagcgtg ag #aagatcaa   2580 gcagttctcc cagcaggagg agaagaggca gaagtcggag cggctgcagc aa #cagcagaa   2640 acacgagaac cagatgcggg acatgctggc gcagtgcgag agcaacatga gc #gagctgca   2700 gcagctgcag aatgaaaagt gccacctcct ggtagagcac gaaacccaga aa #ctgaaggc   2760 cctggatgag agccataacc agaacctgaa ggaatggcgg gacaagcttc gg #ccgcgcaa   2820 gaaggctctg gaagaggatc tgaaccagaa gaagcgggag caggagatgt tc #ttcaagct   2880 gagcgaggag gcggagtgcc caaacccctc caccccaagc aaggccgcca ag #ttcttccc   2940 ctacagctct ggggatgctt cc            #                   #               2962 <210> SEQ ID NO 107 <211> LENGTH: 968 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 107 Met Ala Phe Ala Asn Phe Arg Arg Ile Leu Ar #g Leu Ser Thr Phe Glu   1               5  #                 10  #                 15 Lys Arg Lys Ser Arg Glu Tyr Glu His Val Ar #g Arg Asp Leu Asp Pro              20      #             25      #             30 Asn Glu Val Trp Glu Ile Val Gly Glu Leu Gl #y Asp Gly Ala Phe Gly          35          #         40          #         45 Lys Val Tyr Lys Ala Lys Asn Lys Glu Thr Gl #y Ala Leu Ala Ala Ala      50              #     55              #     60 Lys Val Ile Glu Thr Lys Ser Glu Glu Glu Le #u Glu Asp Tyr Ile Val  65                  # 70                  # 75                  # 80 Glu Ile Glu Ile Leu Ala Thr Cys Asp His Pr #o Tyr Ile Val Lys Leu                  85  #                 90  #                 95 Leu Gly Ala Tyr Tyr His Asp Gly Lys Leu Tr #p Ile Met Ile Glu Phe             100       #           105       #           110 Cys Pro Gly Gly Ala Val Asp Ala Ile Met Le #u Glu Leu Asp Arg Gly         115           #       120           #       125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Ar #g Gln Met Leu Glu Ala     130               #   135               #   140 Leu Asn Phe Leu His Ser Lys Arg Ile Ile Hi #s Arg Asp Leu Lys Ala 145                 1 #50                 1 #55                 1 #60 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Il #e Arg Leu Ala Asp Phe                 165   #               170   #               175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gl #n Lys Arg Asp Ser Phe             180       #           185       #           190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Va #l Val Met Cys Glu Thr         195           #       200           #       205 Met Lys Asp Thr Pro Tyr Asp Tyr Lys Ala As #p Ile Trp Ser Leu Gly     210               #   215               #   220 Ile Thr Leu Ile Glu Met Ala Gln Ile Glu Pr #o Pro His His Glu Leu 225                 2 #30                 2 #35                 2 #40 Asn Pro Met Arg Val Leu Leu Lys Ile Ala Ly #s Ser Asp Pro Pro Thr                 245   #               250   #               255 Leu Leu Thr Pro Ser Lys Trp Ser Val Glu Ph #e Arg Asp Phe Leu Lys             260       #           265       #           270 Ile Ala Leu Asp Lys Asn Pro Glu Thr Arg Pr #o Ser Ala Ala Gln Leu         275           #       280           #       285 Leu Glu His Pro Phe Val Ser Ser Ile Thr Se #r Asn Lys Ala Leu Arg     290               #   295               #   300 Glu Leu Val Ala Glu Ala Lys Ala Glu Val Me #t Glu Glu Ile Glu Asp 305                 3 #10                 3 #15                 3 #20 Gly Arg Asp Glu Gly Glu Glu Glu Asp Ala Va #l Asp Ala Ala Ser Thr                 325   #               330   #               335 Leu Glu Asn His Thr Gln Asn Ser Ser Glu Va #l Ser Pro Pro Ser Leu             340       #           345       #           350 Asn Ala Asp Lys Pro Leu Glu Glu Ser Pro Se #r Thr Pro Leu Ala Pro         355           #       360           #       365 Ser Gln Ser Gln Asp Ser Val Asn Glu Pro Cy #s Ser Gln Pro Ser Gly     370               #   375               #   380 Asp Arg Ser Leu Gln Thr Thr Ser Pro Pro Va #l Val Ala Pro Gly Asn 385                 3 #90                 3 #95                 4 #00 Glu Asn Gly Leu Ala Val Pro Val Pro Leu Ar #g Lys Ser Arg Pro Val                 405   #               410   #               415 Ser Met Asp Ala Arg Ile Gln Val Ala Gln Gl #u Lys Gln Val Ala Glu             420       #           425       #           430 Gln Gly Gly Asp Leu Ser Pro Ala Ala Asn Ar #g Ser Gln Lys Ala Ser         435           #       440           #       445 Gln Ser Arg Pro Asn Ser Ser Ala Leu Glu Th #r Leu Gly Gly Glu Lys     450               #   455               #   460 Leu Ala Asn Gly Ser Leu Glu Pro Pro Ala Gl #n Ala Ala Pro Gly Pro 465                 4 #70                 4 #75                 4 #80 Ser Lys Arg Asp Ser Asp Cys Ser Ser Leu Cy #s Thr Ser Glu Ser Met                 485   #               490   #               495 Asp Tyr Gly Thr Asn Leu Ser Thr Asp Leu Se #r Leu Asn Lys Glu Met             500       #           505       #           510 Gly Ser Leu Ser Ile Lys Asp Pro Lys Leu Ty #r Lys Lys Thr Leu Lys         515           #       520           #       525 Arg Thr Arg Lys Phe Val Val Asp Gly Val Gl #u Val Ser Ile Thr Thr     530               #   535               #   540 Ser Lys Ile Ile Ser Glu Asp Glu Lys Lys As #p Glu Glu Met Arg Phe 545                 5 #50                 5 #55                 5 #60 Leu Arg Arg Gln Glu Leu Arg Glu Leu Arg Le #u Leu Gln Lys Glu Glu                 565   #               570   #               575 His Arg Asn Gln Thr Gln Leu Ser Asn Lys Hi #s Glu Leu Gln Leu Glu             580       #           585       #           590 Gln Met His Lys Arg Phe Glu Gln Glu Ile As #n Ala Lys Lys Lys Phe         595           #       600           #       605 Phe Asp Thr Glu Leu Glu Asn Leu Glu Arg Gl #n Gln Lys Gln Gln Val     610               #   615               #   620 Glu Lys Met Glu Gln Asp His Ala Val Arg Ar #g Arg Glu Glu Ala Arg 625                 6 #30                 6 #35                 6 #40 Arg Ile Arg Leu Glu Gln Asp Arg Asp Tyr Th #r Arg Phe Gln Glu Gln                 645   #               650   #               655 Leu Lys Leu Met Lys Lys Glu Val Lys Asn Gl #u Val Glu Lys Leu Pro             660       #           665       #           670 Arg Gln Gln Arg Lys Glu Ser Met Lys Gln Ly #s Met Glu Glu His Thr         675           #       680           #       685 Gln Lys Lys Gln Leu Leu Asp Arg Asp Phe Va #l Ala Lys Gln Lys Glu     690               #   695               #   700 Asp Leu Glu Leu Ala Met Lys Arg Leu Thr Th #r Asp Asn Arg Arg Glu 705                 7 #10                 7 #15                 7 #20 Ile Cys Asp Lys Glu Arg Glu Cys Leu Met Ly #s Lys Gln Glu Leu Leu                 725   #               730   #               735 Arg Asp Arg Glu Ala Ala Leu Trp Glu Met Gl #u Glu His Gln Leu Gln             740       #           745       #           750 Glu Arg His Gln Leu Val Lys Gln Gln Leu Ly #s Asp Gln Tyr Phe Leu         755           #       760           #       765 Gln Arg His Glu Leu Leu Arg Lys His Glu Ly #s Glu Arg Glu Gln Met     770               #   775               #   780 Gln Arg Tyr Asn Gln Arg Met Ile Glu Gln Le #u Lys Val Arg Gln Gln 785                 7 #90                 7 #95                 8 #00 Gln Glu Lys Ala Arg Leu Pro Lys Ile Gln Ar #g Ser Glu Gly Lys Thr                 805   #               810   #               815 Arg Met Ala Met Tyr Lys Lys Ser Leu His Il #e Asn Gly Gly Gly Ser             820       #           825       #           830 Ala Ala Glu Gln Arg Glu Lys Ile Lys Gln Ph #e Ser Gln Gln Glu Glu         835           #       840           #       845 Lys Arg Gln Lys Ser Glu Arg Leu Gln Gln Gl #n Gln Lys His Glu Asn     850               #   855               #   860 Gln Met Arg Asp Met Leu Ala Gln Cys Glu Se #r Asn Met Ser Glu Leu 865                 8 #70                 8 #75                 8 #80 Gln Gln Leu Gln Asn Glu Lys Cys His Leu Le #u Val Glu His Glu Thr                 885   #               890   #               895 Gln Lys Leu Lys Ala Leu Asp Glu Ser His As #n Gln Asn Leu Lys Glu             900       #           905       #           910 Trp Arg Asp Lys Leu Arg Pro Arg Lys Lys Al #a Leu Glu Glu Asp Leu         915           #       920           #       925 Asn Gln Lys Lys Arg Glu Gln Glu Met Phe Ph #e Lys Leu Ser Glu Glu     930               #   935               #   940 Ala Glu Cys Pro Asn Pro Ser Thr Pro Ser Ly #s Ala Ala Lys Phe Phe 945                 9 #50                 9 #55                 9 #60 Pro Tyr Ser Ser Gly Asp Ala Ser                 965 <210> SEQ ID NO 108 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (2) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (4) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (6) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (8) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (10) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 108 Leu Xaa Leu Xaa Leu Xaa Leu Xaa Leu Xaa Le #u   1               5  #                 10 <210> SEQ ID NO 109 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 109 gcagcaagtg gagaagatgg             #                   #                   # 20 <210> SEQ ID NO 110 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 110 ggaagcatcc ccagagctgt ag            #                   #                 22 <210> SEQ ID NO 111 <211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 111 Glu Lys Phe Gln Lys Cys Ser Ala Asp Glu Se #r Pro   1               5  #                 10 <210> SEQ ID NO 112 <211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 112 Ser Ile Ser Asn Ser Glu Leu Phe Pro Thr Th #r Asp Pro Val Gly Thr   1               5  #                 10  #                 15 <210> SEQ ID NO 113 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 113 Leu Asp Phe Pro Lys Glu Asp Tyr Arg   1               5 <210> SEQ ID NO 114 <211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 114 His Gly Asp Pro Arg Pro Glu Pro Arg Pro Th #r Gln   1               5  #                 10 <210> SEQ ID NO 115 <211> LENGTH: 14 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 115 Pro Ser Thr Asn Arg Ala Gly Ser Leu Lys As #p Pro Glu Cys   1               5  #                 10 <210> SEQ ID NO 116 <211> LENGTH: 19 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 116 Asp Pro Arg Thr Arg Ala Ser Asp Pro Gln Se #r Pro Pro Gln Val Ser   1               5  #                 10  #                 15 Arg His Lys <210> SEQ ID NO 117 <211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 117 Cys Leu Val Pro Leu Ile Gln Leu Tyr Arg Ly #s Gln Thr Ser Thr Cys   1               5  #                 10  #                 15 <210> SEQ ID NO 118 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 118 Pro Leu Met Arg Gln Asn Arg Thr Arg  1               5 <210> SEQ ID NO 119 <211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 119 Ser Gly Asp Arg Arg Arg Ala Gly Pro Glu Ly #s Arg Pro Lys Ser Ser   1               5  #                 10  #                 15 <210> SEQ ID NO 120 <211> LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 120 Cys Arg Arg Lys Ser Leu Val Gly Thr Pro Ty #r Trp Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 121 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 121 Arg Thr Val Gly Arg Arg Asn Thr Phe Ile Gl #y Thr Pro Pro Tyr   1               5  #                 10  #                 15 <210> SEQ ID NO 122 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 122 Lys Arg Lys Ser Phe Ile Gly Thr Pro Tyr Tr #p Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 123 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 123 Lys Arg Asn Thr Phe Val Gly Thr Pro Phe Tr #p Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 124 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 124 Pro Ala Asn Ser Phe Val Gly Thr Pro Tyr Tr #p Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 125 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 125 Arg Arg Asn Thr Phe Ile Gly Thr Pro Tyr Tr #p Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 126 <211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 126 Arg Arg Asn Thr Phe Ile Gly Thr Pro Tyr Tr #p Met Ala Pro Glu   1               5  #                 10  #                 15 <210> SEQ ID NO 127 <211> LENGTH: 18 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 127 Arg Asn Lys Val Arg Lys Thr Phe Val Gly Th #r Pro Cys Trp Met Ala   1               5  #                 10  #                 15 Pro Glu <210> SEQ ID NO 128 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 128 ctcccatttc ctagcaaaat ca            #                   #                 22 <210> SEQ ID NO 129 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 129 agaggcagta ttgtcagatg ta            #                   #                 22 <210> SEQ ID NO 130 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 130 ccacacatgc gtatctctgt tg            #                   #                 22 <210> SEQ ID NO 131 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 131 ttgctagaat tcacatcagg taca           #                   #                24 <210> SEQ ID NO 132 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 132 atccctggat cacactgctt ct            #                   #                 22 <210> SEQ ID NO 133 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 133 caaggtgttc tttgcctctg tt            #                   #                 22 <210> SEQ ID NO 134 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 134 agatggactg tactgggagg g            #                   #                   #21 <210> SEQ ID NO 135 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 135 agaagagcac ttggcactta tc            #                   #                 22 <210> SEQ ID NO 136 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 136 catcatgaac tggtgacggg             #                   #                   # 20 <210> SEQ ID NO 137 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 137 ccagtgaaat caaaccagta aaa            #                   #                23 <210> SEQ ID NO 138 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 138 caaaacctgg ccgtctcttc tatt           #                   #                24 <210> SEQ ID NO 139 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 139 atttgtgcta ctgggattct gtg            #                   #                23 <210> SEQ ID NO 140 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 140 gaatagcggt accatgatag aata           #                   #                24 <210> SEQ ID NO 141 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 141 taccaaaaag agccaaaagt gtg            #                   #                23 <210> SEQ ID NO 142 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 142 ctcagtattc tctccaaaga ttg            #                   #                23 <210> SEQ ID NO 143 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 143 gatgttctct ccattctgta aag            #                   #                23 <210> SEQ ID NO 144 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 144 catcactgga agtctgcagt g            #                   #                   #21 <210> SEQ ID NO 145 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <400> SEQUENCE: 145 caggtgcagt agtcatttgc             #                   #                   # 20 <210> SEQ ID NO 146 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 146 ggagctgtcg tattccagtc             #                   #                   # 20 <210> SEQ ID NO 147 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Primer <400> SEQUENCE: 147 aacccctcaa gacccgttta g            #                   #                   #21 <210> SEQ ID NO 148 <211> LENGTH: 4 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       motif <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (2) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (3) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 148 Pro Xaa Xaa Pro   1 <210> SEQ ID NO 149 <211> LENGTH: 50 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       leucine zipper <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (2)..(7) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (9)..(14) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (16)..(21) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (23)..(42) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (44)..(49) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 149 Leu Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Xa #a Xaa Xaa Xaa Leu Xaa   1               5  #                 10  #                 15 Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Xa #a Xaa Xaa Xaa Xaa Xaa              20      #             25      #             30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Le #u Xaa Xaa Xaa Xaa Xaa          35          #         40          #         45 Xaa Leu      50 <210> SEQ ID NO 150 <211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Description of Artificial  #Sequence: Synthetic       Cdc42/Rac-binding motif <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (3)..(5) <223> OTHER INFORMATION: Any amino acid <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (7)..(8) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 150 Ser Pro Xaa Xaa Xaa His Xaa Xaa His   1               5 <210> SEQ ID NO 151 <211> LENGTH: 787 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 151 Ile Ile Glu Leu Ala Glu Arg Lys Pro Pro Le #u Phe Asn Met Asn Ala   1               5  #                 10  #                 15 Met Ser Ala Leu Tyr His Ile Ala Gln Asn Gl #u Ser Pro Thr Leu Gln              20      #             25      #             30 Ser Asn Glu Trp Ser Asp Tyr Phe Arg Asn Ph #e Val Asp Ser Cys Leu          35          #         40          #         45 Gln Lys Ile Pro Gln Asp Arg Pro Thr Ser Gl #u Glu Leu Leu Lys His      50              #     55              #     60 Ile Phe Val Leu Arg Glu Arg Pro Glu Thr Va #l Leu Ile Asp Leu Ile  65                  # 70                  # 75                  # 80 Gln Arg Thr Lys Asp Ala Val Arg Glu Leu As #p Asn Leu Gln Tyr Arg                  85  #                 90  #                 95 Lys Met Lys Lys Leu Leu Phe Gln Glu Ala Hi #s Asn Gly Pro Ala Tyr             100       #           105       #           110 Glu Ala Gln Glu Glu Glu Glu Glu Gln Asp Hi #s Gly Val Gly Arg Thr         115           #       120           #       125 Gly Thr Val Asn Ser Val Gly Ser Asn Gln Se #r Ile Pro Ser Met Ser     130               #   135               #   140 Ile Ser Ala Ser Ser Gln Ser Ser Ser Val As #n Ser Leu Pro Asp Val 145                 1 #50                 1 #55                 1 #60 Ser Asp Asp Lys Ser Glu Leu Asp Met Met Gl #u Gly Asp His Thr Val                 165   #               170   #               175 Met Ser Asn Ser Ser Val Ile His Leu Lys Pr #o Glu Glu Glu Asn Tyr             180       #           185       #           190 Arg Glu Glu Gly Asp Pro Arg Thr Arg Ala Se #r Asp Pro Gln Ser Pro         195           #       200           #       205 Pro Gln Val Ser Arg His Lys Ser His Tyr Ar #g Asn Arg Glu His Phe     210               #   215               #   220 Ala Thr Ile Arg Thr Ala Ser Leu Val Thr Ar #g Gln Met Gln Glu His 225                 2 #30                 2 #35                 2 #40 Glu Gln Asp Ser Glu Leu Arg Glu Gln Met Se #r Gly Tyr Lys Arg Met                 245   #               250   #               255 Arg Arg Gln His Gln Lys Gln Leu Met Thr Le #u Glu Asn Lys Leu Lys             260       #           265       #           270 Ala Glu Met Asp Glu His Arg Leu Arg Leu As #p Lys Cys Leu Glu Thr         275           #       280           #       285 Gly Arg Asn Asn Phe Ala Ala Glu Met Glu Ly #s Leu Ile Lys Lys His     290               #   295               #   300 Gln Ala Ala Met Glu Lys Glu Ala Lys Val Me #t Ser Asn Glu Glu Lys 305                 3 #10                 3 #15                 3 #20 Lys Phe Gln Gln His Ile Gln Ala Gln Gln Ly #s Lys Glu Leu Asn Ser                 325   #               330   #               335 Phe Leu Glu Ser Gln Lys Arg Glu Tyr Lys Le #u Arg Lys Glu Gln Leu             340       #           345       #           350 Lys Glu Glu Leu Asn Glu Asn Gln Ser Thr Pr #o Lys Lys Glu Lys Gln         355           #       360           #       365 Glu Trp Leu Ser Lys Gln Lys Glu Asn Ile Gl #n His Phe Gln Ala Glu     370               #   375               #   380 Glu Glu Ala Asn Leu Leu Arg Arg Gln Arg Gl #n Tyr Leu Glu Leu Glu 385                 3 #90                 3 #95                 4 #00 Cys Arg Arg Phe Lys Arg Arg Met Leu Leu Gl #y Arg His Asn Leu Glu                 405   #               410   #               415 Gln Asp Leu Val Arg Glu Glu Leu Asn Lys Ar #g Gln Thr Gln Lys Asp             420       #           425       #           430 Leu Glu His Ala Met Leu Leu Arg Gln His Gl #u Ser Met Gln Glu Leu         435           #       440           #       445 Glu Phe Arg His Leu Asn Thr Ile Gln Lys Me #t Arg Cys Glu Leu Ile     450               #   455               #   460 Arg Leu Gln His Gln Thr Glu Leu Thr Asn Gl #n Leu Glu Tyr Asn Lys 465                 4 #70                 4 #75                 4 #80 Arg Arg Glu Arg Glu Leu Arg Arg Lys His Va #l Met Glu Val Arg Gln                 485   #               490   #               495 Gln Pro Lys Ser Leu Lys Ser Lys Glu Leu Gl #n Ile Lys Lys Gln Phe             500       #           505       #           510 Gln Asp Thr Cys Lys Ile Gln Thr Arg Gln Ty #r Lys Ala Leu Arg Asn         515           #       520           #       525 His Leu Leu Glu Thr Thr Pro Lys Ser Glu Hi #s Lys Ala Val Leu Lys     530               #   535               #   540 Arg Leu Lys Glu Glu Gln Thr Arg Lys Leu Al #a Ile Leu Ala Glu Gln 545                 5 #50                 5 #55                 5 #60 Tyr Asp His Ser Ile Asn Glu Met Leu Ser Th #r Gln Ala Leu Arg Leu                 565   #               570   #               575 Asp Glu Ala Gln Glu Ala Glu Cys Gln Val Le #u Lys Met Gln Leu Gln             580       #           585       #           590 Gln Glu Leu Glu Leu Leu Asn Ala Tyr Gln Se #r Lys Ile Lys Met Gln         595           #       600           #       605 Ala Glu Ala Gln His Asp Arg Glu Leu Arg Gl #u Leu Glu Gln Arg Val     610               #   615               #   620 Ser Leu Arg Arg Ala Leu Leu Glu Gln Lys Il #e Glu Glu Glu Met Leu 625                 6 #30                 6 #35                 6 #40 Ala Leu Gln Asn Glu Arg Thr Glu Arg Ile Ar #g Ser Leu Leu Glu Arg                 645   #               650   #               655 Gln Ala Arg Glu Ile Glu Ala Phe Asp Ser Gl #u Ser Met Arg Leu Gly             660       #           665       #           670 Phe Ser Asn Met Val Leu Ser Asn Leu Ser Pr #o Glu Ala Phe Ser His         675           #       680           #       685 Ser Tyr Pro Gly Ala Ser Gly Trp Ser His As #n Pro Thr Gly Gly Pro     690               #   695               #   700 Gly Pro His Trp Gly His Pro Met Gly Gly Pr #o Pro Gln Ala Trp Gly 705                 7 #10                 7 #15                 7 #20 His Pro Met Gln Gly Gly Pro Gln Pro Trp Gl #y His Pro Ser Gly Pro                 725   #               730   #               735 Met Gln Gly Val Pro Arg Gly Ser Ser Met Gl #y Val Arg Asn Ser Pro             740       #           745       #           750 Gln Ala Leu Arg Arg Thr Ala Ser Gly Gly Ar #g Thr Glu Gln Gly Met         755           #       760           #       765 Ser Arg Ser Thr Ser Val Thr Ser Gln Ile Se #r Asn Gly Ser His Met     770               #   775               #   780 Ser Tyr Thr 785 <210> SEQ ID NO 152 <211> LENGTH: 338 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION: (2) <223> OTHER INFORMATION: Any amino acid <400> SEQUENCE: 152 His Xaa Glu Tyr Val Pro Val Arg Arg Ile As #n Leu Glu Ala Cys Ser   1               5  #                 10  #                 15 Asn Glu Met Val Thr Ser Cys Arg Ala Ser Cy #s Met Phe Gln Thr Leu              20      #             25      #             30 Asn His Pro Asn Ile Val Pro Tyr Arg Ala Th #r Leu Ile Ala Asp Asn          35          #         40          #         45 Glu Leu Trp Val Val Thr Ser Phe Met Ala Ty #r Gly Ser Ala Lys Asp      50              #     55              #     60 Leu Ile Cys Thr His Phe Met Asp Gly Met As #n Glu Leu Ala Ile Ala  65                  # 70                  # 75                  # 80 Tyr Ile Leu Gln Gly Val Leu Lys Ala Leu As #p Tyr Ile His His Asn                  85  #                 90  #                 95 Gly Tyr Val His Arg Ser Val Lys Ala Ser Hi #s Ile Leu Ile Ser Val             100       #           105       #           110 Asp Gly Lys Val Tyr Leu Ser Gly Leu Arg Se #r Asn Leu Ser Met Ile         115           #       120           #       125 Ser His Gly Gln Arg Gln Arg Val Val His As #p Phe Pro Lys Tyr Ser     130               #   135               #   140 Val Lys Val Leu Pro Trp Leu Ser Pro Glu Va #l Leu Gln Gln Asn Leu 145                 1 #50                 1 #55                 1 #60 Gln Gly Tyr Asp Ala Lys Ser Asp Ile Tyr Se #r Val Gly Ile Thr Ala                 165   #               170   #               175 Cys Glu Leu Ala Asn Gly His Val Pro Phe Ly #s Asp Met Pro Ala Thr             180       #           185       #           190 Gln Met Leu Leu Glu Lys Leu Asn Gly Thr Va #l Pro Cys Leu Leu Asp         195           #       200           #       205 Thr Ser Thr Ile Pro Ala Glu Glu Leu Thr Me #t Ser Pro Ser Arg Ser     210               #   215               #   220 Val Ala Asn Ser Gly Leu Ser Asp Ser Leu Th #r Thr Ser Thr Pro Arg 225                 2 #30                 2 #35                 2 #40 Pro Ser Asn Gly Asp Ser Pro Ser His Pro Ty #r His Arg Thr Phe Ser                 245   #               250   #               255 Pro His Phe His His Phe Val Glu Gln Cys Le #u Gln Arg Asn Pro Asp             260       #           265       #           270 Ala Arg Pro Ser Ala Ser Thr Leu Leu Asn Hi #s Ser Phe Phe Lys Gln         275           #       280           #       285 Ile Lys Arg Arg Ala Ser Glu Ala Leu Pro Gl #u Leu Leu Arg Pro Val     290               #   295               #   300 Thr Pro Ile Thr Asn Phe Glu Gly Ser Gln Se #r Gln Asp His Ser Gly 305                 3 #10                 3 #15                 3 #20 Ile Phe Gly Leu Val Thr Asn Leu Glu Glu Le #u Glu Val Asp Asp Trp                 325   #               330   #               335 Glu Phe <210> SEQ ID NO 153 <211> LENGTH: 546 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 153 Met Ala Glu Pro Ser Gly Ser Val His Val Gl #n Leu Pro Gln Gln Ala   1               5  #                 10  #                 15 Ala Pro Val Thr Ala Ala Ala Ala Ala Ala Pr #o Ala Ala Ala Thr Ala              20      #             25      #             30 Ala Pro Ala Pro Ala Ala Pro Ala Ala Pro Al #a Pro Ala Pro Ala Pro          35          #         40          #         45 Ala Pro Ala Ala Gln Ala Val Gly Trp Pro Il #e Cys Arg Asp Ala Tyr      50              #     55              #     60 Glu Leu Gln Glu Val Ile Gly Ser Gly Ala Th #r Ala Val Val Gln Ala  65                  # 70                  # 75                  # 80 Ala Leu Cys Lys Pro Arg Gln Glu Arg Val Al #a Ile Lys Arg Ile Asn                  85  #                 90  #                 95 Leu Glu Lys Cys Gln Thr Ser Met Asp Glu Le #u Leu Lys Glu Ile Gln             100       #           105       #           110 Ala Met Ser Gln Cys Ser His Pro Asn Val Va #l Thr Tyr Tyr Thr Ser         115           #       120           #       125 Phe Val Val Lys Asp Glu Leu Trp Leu Val Me #t Lys Leu Leu Ser Gly     130               #   135               #   140 Gly Ser Met Leu Asp Ile Ile Lys Tyr Ile Va #l Asn Arg Gly Glu His 145                 1 #50                 1 #55                 1 #60 Lys Asn Gly Val Leu Glu Glu Ala Ile Ile Al #a Thr Ile Leu Lys Glu                 165   #               170   #               175 Val Leu Glu Gly Leu Asp Tyr Leu His Arg As #n Gly Gln Ile His Arg             180       #           185       #           190 Asp Leu Lys Ala Gly Asn Ile Leu Leu Gly Gl #u Asp Gly Ser Val Gln         195           #       200           #       205 Ile Ala Asp Phe Gly Val Ser Ala Phe Leu Al #a Thr Gly Gly Asp Val     210               #   215               #   220 Thr Arg Asn Lys Val Arg Lys Thr Phe Val Gl #y Thr Pro Cys Trp Met 225                 2 #30                 2 #35                 2 #40 Ala Pro Glu Val Met Glu Gln Val Arg Gly Ty #r Asp Phe Lys Ala Asp                 245   #               250   #               255 Met Trp Ser Phe Gly Ile Thr Ala Ile Glu Le #u Ala Thr Gly Ala Ala             260       #           265       #           270 Pro Tyr His Lys Tyr Pro Pro Met Lys Val Le #u Met Leu Thr Leu Gln         275           #       280           #       285 Asn Asp Pro Pro Thr Leu Glu Thr Gly Val Gl #u Asp Lys Glu Met Met     290               #   295               #   300 Lys Lys Tyr Gly Lys Ser Phe Arg Lys Leu Le #u Ser Leu Cys Leu Gln 305                 3 #10                 3 #15                 3 #20 Lys Asp Pro Ser Lys Arg Pro Thr Ala Ala Gl #u Leu Leu Lys Cys Lys                 325   #               330   #               335 Phe Phe Gln Lys Ala Lys Asn Arg Glu Tyr Le #u Ile Glu Lys Leu Leu             340       #           345       #           350 Thr Arg Thr Pro Asp Ile Ala Gln Arg Ala Ly #s Lys Val Arg Arg Val         355           #       360           #       365 Pro Gly Ser Ser Gly His Leu His Lys Thr Gl #u Asp Gly Asp Trp Glu     370               #   375               #   380 Trp Ser Asp Asp Glu Met Asp Glu Lys Ser Gl #u Glu Gly Lys Ala Ala 385                 3 #90                 3 #95                 4 #00 Phe Ser Gln Glu Lys Ser Arg Arg Val Lys Gl #u Glu Asn Pro Glu Ile                 405   #               410   #               415 Ala Val Ser Ala Ser Thr Ile Pro Glu Gln Il #e Gln Ser Leu Ser Val             420       #           425       #           430 His Asp Ser Gln Gly Pro Pro Asn Ala Asn Gl #u Asp Tyr Arg Glu Ala         435           #       440           #       445 Ser Ser Cys Ala Val Asn Leu Val Leu Arg Le #u Arg Asn Ser Arg Lys     450               #   455               #   460 Glu Leu Asn Asp Ile Arg Phe Glu Phe Thr Pr #o Gly Arg Asp Thr Ala 465                 4 #70                 4 #75                 4 #80 Asp Gly Val Ser Gln Glu Leu Phe Ser Ala Gl #y Leu Val Asp Gly His                 485   #               490   #               495 Asp Val Val Ile Val Ala Ala Asn Leu Gln Ly #s Ile Val Asp Asp Pro             500       #           505       #           510 Lys Ala Leu Lys Thr Leu Thr Phe Lys Leu Al #a Ser Gly Cys Asp Gly         515           #       520           #       525 Ser Glu Ile Pro Asp Glu Val Lys Leu Ile Gl #y Phe Ala Gln Leu Ser     530               #   535               #   540 Val Ser 545 <210> SEQ ID NO 154 <211> LENGTH: 966 <212> TYPE: PRT <213> ORGANISM: Murine sp. <400> SEQUENCE: 154 Met Ala Phe Ala Asn Phe Arg Arg Ile Leu Ar #g Leu Ser Thr Phe Glu   1               5  #                 10  #                 15 Lys Arg Lys Ser Arg Glu Tyr Glu His Val Ar #g Arg Asp Leu Asp Pro              20      #             25      #             30 Asn Asp Val Trp Glu Ile Val Gly Glu Leu Gl #y Asp Gly Ala Phe Gly          35          #         40          #         45 Lys Val Tyr Lys Ala Lys Asn Lys Glu Thr Gl #y Ala Leu Ala Ala Ala      50              #     55              #     60 Lys Val Ile Glu Thr Lys Ser Glu Glu Glu Le #u Glu Asp Tyr Ile Val  65                  # 70                  # 75                  # 80 Glu Ile Glu Ile Leu Ala Thr Cys Asp His Pr #o Tyr Ile Val Lys Leu                  85  #                 90  #                 95 Leu Gly Ala Tyr Tyr Tyr Asp Gly Lys Leu Tr #p Ile Met Ile Glu Phe             100       #           105       #           110 Cys Pro Gly Gly Ala Val Asp Ala Ile Met Le #u Glu Leu Asp Arg Gly         115           #       120           #       125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Ar #g Gln Met Leu Glu Ala     130               #   135               #   140 Leu Asn Phe Leu His Gly Lys Arg Ile Ile Hi #s Arg Asp Leu Lys Ala 145                 1 #50                 1 #55                 1 #60 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Il #e Arg Leu Ala Asp Phe                 165   #               170   #               175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gl #n Lys Arg Asp Ser Phe             180       #           185       #           190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Va #l Val Leu Cys Glu Thr         195           #       200           #       205 Met Lys Asp Ala Pro Tyr Asp Tyr Lys Ala As #p Ile Trp Ser Leu Gly     210               #   215               #   220 Ile Thr Leu Ile Glu Met Ala Gln Ile Glu Pr #o Pro His His Glu Leu 225                 2 #30                 2 #35                 2 #40 Asn Pro Met Arg Val Leu Leu Lys Ile Ala Ly #s Ser Asp Pro Pro Thr                 245   #               250   #               255 Leu Leu Thr Pro Ser Lys Trp Ser Val Glu Ph #e Arg Asp Phe Leu Lys             260       #           265       #           270 Ile Ala Leu Asp Lys Asn Pro Glu Thr Arg Pr #o Ser Ala Ala Gln Leu         275           #       280           #       285 Leu Gln His Pro Phe Val Ser Arg Val Thr Se #r Asn Lys Ala Leu Arg     290               #   295               #   300 Glu Leu Val Ala Glu Ala Lys Ala Glu Val Me #t Glu Glu Ile Glu Asp 305                 3 #10                 3 #15                 3 #20 Gly Arg Glu Asp Gly Glu Glu Glu Asp Ala Va #l Asp Ala Val Pro Pro                 325   #               330   #               335 Leu Val Asn His Thr Gln Asp Ser Ala Asn Va #l Thr Gln Pro Ser Leu             340       #           345       #           350 Asp Ser Asn Lys Leu Leu Gln Asp Ser Ser Th #r Pro Leu Pro Pro Ser         355           #       360           #       365 Gln Pro Gln Glu Pro Val Asn Gly Pro Cys Se #r Gln Pro Ser Gly Asp     370               #   375               #   380 Gly Pro Leu Gln Thr Thr Ser Pro Ala Asp Gl #y Leu Ser Lys Asn Asp 385                 3 #90                 3 #95                 4 #00 Asn Asp Leu Lys Val Pro Val Pro Leu Arg Ly #s Ser Arg Pro Leu Ser                 405   #               410   #               415 Met Asp Ala Arg Ile Gln Met Asp Glu Glu Ly #s Gln Ile Pro Asp Gln             420       #           425       #           430 Asp Glu Asn Pro Ser Pro Ala Ala Ser Lys Se #r Gln Lys Ala Asn Gln         435           #       440           #       445 Ser Arg Pro Asn Ser Ser Ala Leu Glu Thr Le #u Gly Gly Glu Ala Leu     450               #   455               #   460 Thr Asn Gly Gly Leu Glu Leu Pro Ser Ser Va #l Thr Pro Ser His Ser 465                 4 #70                 4 #75                 4 #80 Lys Arg Ala Ser Asp Cys Ser Asn Leu Ser Th #r Ser Glu Ser Met Asp                 485   #               490   #               495 Tyr Gly Thr Ser Leu Ser Ala Asp Leu Ser Le #u Asn Lys Glu Thr Gly             500       #           505       #           510 Ser Leu Ser Leu Lys Gly Ser Lys Leu His As #n Lys Thr Leu Lys Arg         515           #       520           #       525 Thr Arg Arg Phe Val Val Asp Gly Val Glu Va #l Ser Ile Thr Thr Ser     530               #   535               #   540 Lys Ile Ile Ser Glu Asp Glu Lys Lys Asp Gl #u Glu Met Arg Phe Leu 545                 5 #50                 5 #55                 5 #60 Arg Arg Gln Glu Leu Arg Glu Leu Arg Leu Le #u Gln Lys Glu Glu His                 565   #               570   #               575 Arg Asn Gln Thr Gln Leu Ser Ser Lys His Gl #u Leu Gln Leu Glu Gln             580       #           585       #           590 Met His Lys Arg Phe Glu Gln Glu Ile Asn Al #a Lys Lys Lys Phe Tyr         595           #       600           #       605 Asp Val Glu Leu Glu Asn Leu Glu Arg Gln Gl #n Lys Gln Gln Val Glu     610               #   615               #   620 Lys Met Glu Gln Asp His Ser Val Arg Arg Ly #s Glu Glu Ala Lys Arg 625                 6 #30                 6 #35                 6 #40 Ile Arg Leu Glu Gln Asp Arg Asp Tyr Ala Ly #s Phe Gln Glu Gln Leu                 645   #               650   #               655 Lys Gln Met Lys Lys Glu Val Lys Ser Glu Va #l Glu Lys Leu Pro Arg             660       #           665       #           670 Gln Gln Arg Lys Glu Ser Met Lys Gln Lys Me #t Glu Glu His Ser Gln         675           #       680           #       685 Lys Lys Gln Arg Leu Asp Arg Asp Phe Val Al #a Lys Gln Lys Glu Asp     690               #   695               #   700 Leu Glu Leu Ala Met Arg Lys Leu Thr Thr Gl #u Asn Arg Arg Glu Ile 705                 7 #10                 7 #15                 7 #20 Cys Asp Lys Glu Arg Asp Cys Leu Ser Lys Ly #s Gln Glu Leu Leu Arg                 725   #               730   #               735 Asp Arg Glu Ala Ala Leu Trp Glu Met Glu Gl #u His Gln Leu Gln Glu             740       #           745       #           750 Arg His Gln Leu Val Lys Gln Gln Leu Lys As #p Gln Tyr Phe Leu Gln         755           #       760           #       765 Arg His Asp Leu Leu Arg Lys His Glu Lys Gl #u Arg Glu Gln Met Gln     770               #   775               #   780 Arg Tyr Asn Gln Arg Met Met Glu Gln Leu Ly #s Val Arg Gln Gln Gln 785                 7 #90                 7 #95                 8 #00 Glu Lys Ala Arg Leu Pro Lys Ile Gln Arg Se #r Asp Gly Glu Thr Arg                 805   #               810   #               815 Met Ala Met Tyr Lys Lys Ser Leu His Ile As #n Gly Ala Gly Ser Ala             820       #           825       #           830 Ser Glu Gln Arg Glu Lys Ile Lys Gln Phe Se #r Gln Gln Glu Glu Lys         835           #       840           #       845 Arg Gln Lys Ala Glu Arg Leu Gln Gln Gln Gl #n Lys His Glu His Gln     850               #   855               #   860 Met Arg Asp Met Val Ala Gln Cys Glu Ser As #n Met Ser Glu Leu Gln 865                 8 #70                 8 #75                 8 #80 Gln Leu Gln Asn Glu Lys Cys Tyr Leu Leu Va #l Glu His Glu Thr Gln                 885   #               890   #               895 Lys Leu Lys Ala Leu Asp Glu Ser His Asn Gl #n Ser Leu Lys Glu Trp             900       #           905       #           910 Arg Asp Lys Leu Arg Pro Arg Lys Lys Ala Le #u Glu Glu Asp Leu Asn         915           #       920           #       925 Gln Lys Lys Arg Glu Gln Glu Met Phe Phe Ly #s Leu Ser Glu Glu Ala     930               #   935               #   940 Glu Pro Arg Pro Thr Thr Pro Ser Lys Ala Se #r Asn Phe Phe Pro Tyr 945                 9 #50                 9 #55                 9 #60 Ser Ser Gly Asp Ala Ser                 965 <210> SEQ ID NO 155 <211> LENGTH: 968 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 155 Met Ala Phe Ala Asn Phe Arg Arg Ile Leu Ar #g Leu Ser Thr Phe Glu   1               5  #                 10  #                 15 Lys Arg Lys Ser Arg Glu Tyr Glu His Val Ar #g Arg Asp Leu Asp Pro              20      #             25      #             30 Asn Glu Val Trp Glu Ile Val Gly Glu Leu Gl #y Asp Gly Ala Phe Gly          35          #         40          #         45 Lys Val Tyr Lys Ala Lys Asn Lys Glu Thr Gl #y Ala Leu Ala Ala Ala      50              #     55              #     60 Lys Val Ile Glu Thr Lys Ser Glu Glu Glu Le #u Glu Asp Tyr Ile Val  65                  # 70                  # 75                  # 80 Glu Ile Glu Ile Leu Ala Thr Cys Asp His Pr #o Tyr Ile Val Lys Leu                  85  #                 90  #                 95 Leu Gly Ala Tyr Tyr His Asp Gly Lys Leu Tr #p Ile Met Ile Glu Phe             100       #           105       #           110 Cys Pro Gly Gly Ala Val Asp Ala Ile Met Le #u Glu Leu Asp Arg Gly         115           #       120           #       125 Leu Thr Glu Pro Gln Ile Gln Val Val Cys Ar #g Gln Met Leu Glu Ala     130               #   135               #   140 Leu Asn Phe Leu His Ser Lys Arg Ile Ile Hi #s Arg Asp Leu Lys Ala 145                 1 #50                 1 #55                 1 #60 Gly Asn Val Leu Met Thr Leu Glu Gly Asp Il #e Arg Leu Ala Asp Phe                 165   #               170   #               175 Gly Val Ser Ala Lys Asn Leu Lys Thr Leu Gl #n Lys Arg Asp Ser Phe             180       #           185       #           190 Ile Gly Thr Pro Tyr Trp Met Ala Pro Glu Va #l Val Met Cys Glu Thr         195           #       200           #       205 Met Lys Asp Thr Pro Tyr Asp Tyr Lys Ala As #p Ile Trp Ser Leu Gly     210               #   215               #   220 Ile Thr Leu Ile Glu Met Ala Gln Ile Glu Pr #o Pro His His Glu Leu 225                 2 #30                 2 #35                 2 #40 Asn Pro Met Arg Val Leu Leu Lys Ile Ala Ly #s Ser Asp Pro Pro Thr                 245   #               250   #               255 Leu Leu Thr Pro Ser Lys Trp Ser Val Glu Ph #e Arg Asp Phe Leu Lys             260       #           265       #           270 Ile Ala Leu Asp Lys Asn Pro Glu Thr Arg Pr #o Ser Ala Ala Gln Leu         275           #       280           #       285 Leu Glu His Pro Phe Val Ser Ser Ile Thr Se #r Asn Lys Ala Leu Arg     290               #   295               #   300 Glu Leu Val Ala Glu Ala Lys Ala Glu Val Me #t Glu Glu Ile Glu Asp 305                 3 #10                 3 #15                 3 #20 Gly Arg Asp Glu Gly Glu Glu Glu Asp Ala Va #l Asp Ala Ala Ser Thr                 325   #               330   #               335 Leu Glu Asn His Thr Gln Asn Ser Ser Glu Va #l Ser Pro Pro Ser Leu             340       #           345       #           350 Asn Ala Asp Lys Pro Leu Glu Glu Ser Pro Se #r Thr Pro Leu Ala Pro         355           #       360           #       365 Ser Gln Ser Gln Asp Ser Val Asn Glu Pro Cy #s Ser Gln Pro Ser Gly     370               #   375               #   380 Asp Arg Ser Leu Gln Thr Thr Ser Pro Pro Va #l Val Ala Pro Gly Asn 385                 3 #90                 3 #95                 4 #00 Glu Asn Gly Leu Ala Val Pro Val Pro Leu Ar #g Lys Ser Arg Pro Val                 405   #               410   #               415 Ser Met Asp Ala Arg Ile Gln Val Ala Gln Gl #u Lys Gln Val Ala Glu             420       #           425       #           430 Gln Gly Gly Asp Leu Ser Pro Ala Ala Asn Ar #g Ser Gln Lys Ala Ser         435           #       440           #       445 Gln Ser Arg Pro Asn Ser Ser Ala Leu Glu Th #r Leu Gly Gly Glu Lys     450               #   455               #   460 Leu Ala Asn Gly Ser Leu Glu Pro Pro Ala Gl #n Ala Ala Pro Gly Pro 465                 4 #70                 4 #75                 4 #80 Ser Lys Arg Asp Ser Asp Cys Ser Ser Leu Cy #s Thr Ser Glu Ser Met                 485   #               490   #               495 Asp Tyr Gly Thr Asn Leu Ser Thr Asp Leu Se #r Leu Asn Lys Glu Met             500       #           505       #           510 Gly Ser Leu Ser Ile Lys Asp Pro Lys Leu Ty #r Lys Lys Thr Leu Lys         515           #       520           #       525 Arg Thr Arg Lys Phe Val Val Asp Gly Val Gl #u Val Ser Ile Thr Thr     530               #   535               #   540 Ser Lys Ile Ile Ser Glu Asp Glu Lys Lys As #p Glu Glu Met Arg Phe 545                 5 #50                 5 #55                 5 #60 Leu Arg Arg Gln Glu Leu Arg Glu Leu Arg Le #u Leu Gln Lys Glu Glu                 565   #               570   #               575 His Arg Asn Gln Thr Gln Leu Ser Asn Lys Hi #s Glu Leu Gln Leu Glu             580       #           585       #           590 Gln Met His Lys Arg Phe Glu Gln Glu Ile As #n Ala Lys Lys Lys Phe         595           #       600           #       605 Phe Asp Thr Glu Leu Glu Asn Leu Glu Arg Gl #n Gln Lys Gln Gln Val     610               #   615               #   620 Glu Lys Met Glu Gln Asp His Ala Val Arg Ar #g Arg Glu Glu Ala Arg 625                 6 #30                 6 #35                 6 #40 Arg Ile Arg Leu Glu Gln Asp Arg Asp Tyr Th #r Arg Phe Gln Glu Gln                 645   #               650   #               655 Leu Lys Leu Met Lys Lys Glu Val Lys Asn Gl #u Val Glu Lys Leu Pro             660       #           665       #           670 Arg Gln Gln Arg Lys Glu Ser Met Lys Gln Ly #s Met Glu Glu His Thr         675           #       680           #       685 Gln Lys Lys Gln Leu Leu Asp Arg Asp Phe Va #l Ala Lys Gln Lys Glu     690               #   695               #   700 Asp Leu Glu Leu Ala Met Lys Arg Leu Thr Th #r Asp Asn Arg Arg Glu 705                 7 #10                 7 #15                 7 #20 Ile Cys Asp Lys Glu Arg Glu Cys Leu Met Ly #s Lys Gln Glu Leu Leu                 725   #               730   #               735 Arg Asp Arg Glu Ala Ala Leu Trp Glu Met Gl #u Glu His Gln Leu Gln             740       #           745       #           750 Glu Arg His Gln Leu Val Lys Gln Gln Leu Ly #s Asp Gln Tyr Phe Leu         755           #       760           #       765 Gln Arg His Glu Leu Leu Arg Lys His Glu Ly #s Glu Arg Glu Gln Met     770               #   775               #   780 Gln Arg Tyr Asn Gln Arg Met Ile Glu Gln Le #u Lys Val Arg Gln Gln 785                 7 #90                 7 #95                 8 #00 Gln Glu Lys Ala Arg Leu Pro Lys Ile Gln Ar #g Ser Glu Gly Lys Thr                 805   #               810   #               815 Arg Met Ala Met Tyr Lys Lys Ser Leu His Il #e Asn Gly Gly Gly Ser             820       #           825       #           830 Ala Ala Glu Gln Arg Glu Lys Ile Lys Gln Ph #e Ser Gln Gln Glu Glu         835           #       840           #       845 Lys Arg Gln Lys Ser Glu Arg Leu Gln Gln Gl #n Gln Lys His Glu Asn     850               #   855               #   860 Gln Met Arg Asp Met Leu Ala Gln Cys Glu Se #r Asn Met Ser Glu Leu 865                 8 #70                 8 #75                 8 #80 Gln Gln Leu Gln Asn Glu Lys Cys His Leu Le #u Val Glu His Glu Thr                 885   #               890   #               895 Gln Lys Leu Lys Ala Leu Asp Glu Ser His As #n Gln Asn Leu Lys Glu             900       #           905       #           910 Trp Arg Asp Lys Leu Arg Pro Arg Lys Lys Al #a Leu Glu Glu Asp Leu         915           #       920           #       925 Asn Gln Lys Lys Arg Glu Gln Glu Met Phe Ph #e Lys Leu Ser Glu Glu     930               #   935               #   940 Ala Glu Cys Pro Asn Pro Ser Thr Pro Ser Ly #s Ala Ala Lys Phe Phe 945                 9 #50                 9 #55                 9 #60 Pro Tyr Ser Ser Gly Asp Ala Ser                 965 

What is claimed is:
 1. An isolated, enriched or purified nucleic acid molecule comprising a nucleotide sequence that: (a) encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 13; (b) is the complement of the nucleotide sequence of (a); (c) is at least 95% identical to a nucleotide sequence of (a) and encodes a polypeptide having kinase activity; (d) encodes a polypeptide having kinase activity and comprising the amino acid sequence of SEQ ID NO: 13, except that it lacks one or more, but not all, of the following segments of amino acid residues: 1-22, 290-526, 527-640, 641-896, or 897-1239 of SEQ ID NO:13; (e) is the complement of the nucleotide sequence of (d); (f) encodes a polypeptide having kinase activity and comprising the amino acid sequence set forth in amino acid residues 23-289 of SEQ ID NO:13; or (g) is the complement of the nucleotide sequence of (f).
 2. The nucleic acid molecule of claim 1, wherein said nucleic acid sequence comprises SEQ ID NO:9.
 3. The nucleic acid molecule of claim 1, wherein said nucleic acid sequence comprises nucleotides 7-3723 of SEQ ID NO:
 9. 4. The nucleic acid molecule of claim 1, wherein said nucleic acid sequence comprises nucleotides 73-3723 of SEQ ID NO:
 9. 5. An isolated, enriched or purified nucleic acid molecule encoding a polypeptide comprising the amino acid residues 23-289 of SEQ ID NO:13.
 6. A recombinant host cell comprising a nucleic acid molecule of any one of claims 1-5.
 7. A nucleic acid probe comprising a nucleotide sequence, wherein said nucleotide sequence is selected from the group consisting of: (a) a polynucleotide that encodes a polypeptide, wherein said polypeptide consists of a fragment of SEQ ID NO: 13 comprising at least 255 contiguous amino acids; (b) the complement of the polynucleotide of (a); (c) a polynucleotide that encodes a polypeptide, wherein said polypeptide consists of a fragment of SEQ ID NO:13 as set forth in amino acid residues 23-289, 290-526, 527-640, 641-896, or 897-1239; and (d) the complement of the polynucleotide of (c).
 8. The nucleic acid probe of claim 1, wherein said fragment comprises at least 260 contiguous amino acids.
 9. The nucleic acid probe of claim 1, wherein said fragment comprises at least 270 contiguous amino acids.
 10. The nucleic acid probe of claim 7, wherein said fragment comprises at least 275 contiguous amino acids.
 11. The nucleic acid probe of claim 7, wherein said fragment comprises at least 300 contiguous amino acids.
 12. The nucleic acid probe of claim 7, wherein said fragment comprises at least 400 contiguous amino acids.
 13. A recombinant host cell comprising a nucleic acid probe of anyone of claims 7-12.
 14. A vector comprising a nucleic acid molecule or a nucleic acid probe of any one of claims 1-5 or
 7. 15. A method for detection of a kinase nucleic acid molecule in a sample, wherein said method comprises: (a) contacting said sample with the nucleic acid probe of any one of claims 7-12 under conditions wherein said kinase nucleic acid molecule and said probe hybridize; and (b) detecting the presence of the hybridization between said probe and said kinase nucleic acid molecule in said sample.
 16. The method of claim 15, wherein said presence of said hybridization in step (b) is a diagnostic tool for detecting cancer.
 17. A method for detection of a kinase nucleic acid molecule in a sample, wherein said method comprises: (a) comparing a kinase nucleic acid molecule in said sample with a kinase nucleic acid molecule in a control sample by contacting said sample and said control sample with the nucleic acid probe of any one of claims 7-12 under conditions wherein said kinase nucleic acid molecule in said sample and in said control sample hybridize with said probe; and (b) detecting a difference in said hybridization with said probe between the kinase nucleic acid molecule in said sample and in said control sample.
 18. The method of claim 17, wherein said difference in said hybridization in step (b) is a diagnostic tool for the detection of cancer. 